Starting phenix.real_space_refine on Fri Jul 19 09:42:16 2024 by dcliebschner =============================================================================== Processing files: ------------------------------------------------------------------------------- Found model, /net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8jy8_36723/07_2024/8jy8_36723.cif Found real_map, /net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8jy8_36723/07_2024/8jy8_36723.map Processing PHIL parameters: ------------------------------------------------------------------------------- Adding command-line PHIL: ------------------------- refinement.macro_cycles=10 scattering_table=electron resolution=2.45 write_initial_geo_file=False Final processed PHIL parameters: ------------------------------------------------------------------------------- data_manager { real_map_files = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8jy8_36723/07_2024/8jy8_36723.map" default_real_map = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8jy8_36723/07_2024/8jy8_36723.map" model { file = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8jy8_36723/07_2024/8jy8_36723.cif" } default_model = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8jy8_36723/07_2024/8jy8_36723.cif" } resolution = 2.45 write_initial_geo_file = False refinement { macro_cycles = 10 } qi { qm_restraints { package { program = *test } } } Starting job =============================================================================== ------------------------------------------------------------------------------- Citation: ********* Afonine PV, Poon BK, Read RJ, Sobolev OV, Terwilliger TC, Urzhumtsev A, Adams PD. (2018) Real-space refinement in PHENIX for cryo-EM and crystallography. Acta Cryst. D74:531-544. Validating inputs Origin is already at (0, 0, 0), no shifts will be applied ------------------------------------------------------------------------------- Processing inputs ***************** Set random seed Set to: 0 Set model cs if undefined Decide on map wrapping Map wrapping is set to: False Normalize map: mean=0, sd=1 Input map: mean= 0.000 sd= 0.260 Set stop_for_unknowns flag Set to: True Assert model is a single copy model Assert all atoms have isotropic ADPs Construct map_model_manager Extract box with map and model Check model and map are aligned Set scattering table Set to: electron Number of scattering types: 4 Type Number sf(0) Gaussians S 32 5.16 5 C 4988 2.51 5 N 1132 2.21 5 O 1240 1.98 5 sf(0) = scattering factor at diffraction angle 0. Process input model Symmetric amino acids flipped None Time to flip residues: 0.02s Monomer Library directory: "/net/cci-filer2/raid1/xp/phenix/phenix-dev-5388/modules/chem_data/mon_lib" Total number of atoms: 7392 Number of models: 1 Model: "" Number of chains: 8 Chain: "A" Number of atoms: 1823 Number of conformers: 1 Conformer: "" Number of residues, atoms: 242, 1823 Classifications: {'peptide': 242} Link IDs: {'PTRANS': 13, 'TRANS': 228} Chain: "B" Number of atoms: 1823 Number of conformers: 1 Conformer: "" Number of residues, atoms: 242, 1823 Classifications: {'peptide': 242} Link IDs: {'PTRANS': 13, 'TRANS': 228} Chain: "C" Number of atoms: 1823 Number of conformers: 1 Conformer: "" Number of residues, atoms: 242, 1823 Classifications: {'peptide': 242} Link IDs: {'PTRANS': 13, 'TRANS': 228} Chain: "D" Number of atoms: 1823 Number of conformers: 1 Conformer: "" Number of residues, atoms: 242, 1823 Classifications: {'peptide': 242} Link IDs: {'PTRANS': 13, 'TRANS': 228} Chain: "A" Number of atoms: 25 Number of conformers: 1 Conformer: "" Number of residues, atoms: 1, 25 Unusual residues: {'PNT': 1} Classifications: {'undetermined': 1} Chain: "B" Number of atoms: 25 Number of conformers: 1 Conformer: "" Number of residues, atoms: 1, 25 Unusual residues: {'PNT': 1} Classifications: {'undetermined': 1} Chain: "C" Number of atoms: 25 Number of conformers: 1 Conformer: "" Number of residues, atoms: 1, 25 Unusual residues: {'PNT': 1} Classifications: {'undetermined': 1} Chain: "D" Number of atoms: 25 Number of conformers: 1 Conformer: "" Number of residues, atoms: 1, 25 Unusual residues: {'PNT': 1} Classifications: {'undetermined': 1} Time building chain proxies: 4.46, per 1000 atoms: 0.60 Number of scatterers: 7392 At special positions: 0 Unit cell: (92.568, 92.568, 63.84, 90, 90, 90) Space group: P 1 (No. 1) Number of sites at special positions: 0 Number of scattering types: 4 Type Number sf(0) S 32 16.00 O 1240 8.00 N 1132 7.00 C 4988 6.00 sf(0) = scattering factor at diffraction angle 0. Number of disulfides: simple=0, symmetry=0 Automatic linking Parameters for automatic linking Linking & cutoffs Metal : Auto - 3.50 Amino acid : False - 1.90 Carbohydrate : True - 1.99 Ligands : True - 1.99 Small molecules : False - 1.98 Amino acid - RNA/DNA : False Number of custom bonds: simple=0, symmetry=0 Time building additional restraints: 2.96 Conformation dependent library (CDL) restraints added in 1.5 seconds 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. Adding C-beta torsion restraints... Number of C-beta restraints generated: 1680 Finding SS restraints... Secondary structure from input PDB file: 44 helices and 0 sheets defined 74.5% alpha, 0.0% beta 0 base pairs and 0 stacking pairs defined. Time for finding SS restraints: 0.61 Creating SS restraints... Processing helix chain 'A' and resid 71 through 98 Processing helix chain 'A' and resid 103 through 122 removed outlier: 3.678A pdb=" N VAL A 120 " --> pdb=" O MET A 116 " (cutoff:3.500A) Processing helix chain 'A' and resid 130 through 140 removed outlier: 3.508A pdb=" N THR A 134 " --> pdb=" O ASN A 130 " (cutoff:3.500A) removed outlier: 3.703A pdb=" N ALA A 138 " --> pdb=" O THR A 134 " (cutoff:3.500A) removed outlier: 3.589A pdb=" N VAL A 139 " --> pdb=" O VAL A 135 " (cutoff:3.500A) removed outlier: 4.128A pdb=" N PHE A 140 " --> pdb=" O GLY A 136 " (cutoff:3.500A) Processing helix chain 'A' and resid 147 through 170 Processing helix chain 'A' and resid 172 through 177 Processing helix chain 'A' and resid 190 through 195 removed outlier: 4.104A pdb=" N PHE A 194 " --> pdb=" O ILE A 190 " (cutoff:3.500A) Processing helix chain 'A' and resid 204 through 226 removed outlier: 3.659A pdb=" N ILE A 208 " --> pdb=" O ILE A 204 " (cutoff:3.500A) removed outlier: 3.582A pdb=" N THR A 215 " --> pdb=" O GLU A 211 " (cutoff:3.500A) Processing helix chain 'A' and resid 236 through 253 Processing helix chain 'A' and resid 261 through 276 Proline residue: A 268 - end of helix Processing helix chain 'A' and resid 282 through 287 removed outlier: 3.610A pdb=" N TYR A 286 " --> pdb=" O SER A 282 " (cutoff:3.500A) Processing helix chain 'A' and resid 288 through 310 removed outlier: 3.523A pdb=" N LEU A 292 " --> pdb=" O PHE A 288 " (cutoff:3.500A) Proline residue: A 295 - end of helix Processing helix chain 'B' and resid 72 through 98 Processing helix chain 'B' and resid 103 through 122 removed outlier: 3.678A pdb=" N VAL B 120 " --> pdb=" O MET B 116 " (cutoff:3.500A) Processing helix chain 'B' and resid 130 through 140 removed outlier: 3.508A pdb=" N THR B 134 " --> pdb=" O ASN B 130 " (cutoff:3.500A) removed outlier: 3.703A pdb=" N ALA B 138 " --> pdb=" O THR B 134 " (cutoff:3.500A) removed outlier: 3.589A pdb=" N VAL B 139 " --> pdb=" O VAL B 135 " (cutoff:3.500A) removed outlier: 4.128A pdb=" N PHE B 140 " --> pdb=" O GLY B 136 " (cutoff:3.500A) Processing helix chain 'B' and resid 147 through 170 Processing helix chain 'B' and resid 172 through 177 Processing helix chain 'B' and resid 190 through 195 removed outlier: 4.104A pdb=" N PHE B 194 " --> pdb=" O ILE B 190 " (cutoff:3.500A) Processing helix chain 'B' and resid 204 through 226 removed outlier: 3.659A pdb=" N ILE B 208 " --> pdb=" O ILE B 204 " (cutoff:3.500A) removed outlier: 3.582A pdb=" N THR B 215 " --> pdb=" O GLU B 211 " (cutoff:3.500A) Processing helix chain 'B' and resid 236 through 253 Processing helix chain 'B' and resid 261 through 276 Proline residue: B 268 - end of helix Processing helix chain 'B' and resid 282 through 287 removed outlier: 3.610A pdb=" N TYR B 286 " --> pdb=" O SER B 282 " (cutoff:3.500A) Processing helix chain 'B' and resid 288 through 310 removed outlier: 3.523A pdb=" N LEU B 292 " --> pdb=" O PHE B 288 " (cutoff:3.500A) Proline residue: B 295 - end of helix Processing helix chain 'C' and resid 72 through 98 Processing helix chain 'C' and resid 103 through 122 removed outlier: 3.678A pdb=" N VAL C 120 " --> pdb=" O MET C 116 " (cutoff:3.500A) Processing helix chain 'C' and resid 130 through 140 removed outlier: 3.509A pdb=" N THR C 134 " --> pdb=" O ASN C 130 " (cutoff:3.500A) removed outlier: 3.703A pdb=" N ALA C 138 " --> pdb=" O THR C 134 " (cutoff:3.500A) removed outlier: 3.589A pdb=" N VAL C 139 " --> pdb=" O VAL C 135 " (cutoff:3.500A) removed outlier: 4.128A pdb=" N PHE C 140 " --> pdb=" O GLY C 136 " (cutoff:3.500A) Processing helix chain 'C' and resid 147 through 170 Processing helix chain 'C' and resid 172 through 177 Processing helix chain 'C' and resid 190 through 195 removed outlier: 4.104A pdb=" N PHE C 194 " --> pdb=" O ILE C 190 " (cutoff:3.500A) Processing helix chain 'C' and resid 204 through 226 removed outlier: 3.659A pdb=" N ILE C 208 " --> pdb=" O ILE C 204 " (cutoff:3.500A) removed outlier: 3.582A pdb=" N THR C 215 " --> pdb=" O GLU C 211 " (cutoff:3.500A) Processing helix chain 'C' and resid 236 through 253 Processing helix chain 'C' and resid 261 through 276 Proline residue: C 268 - end of helix Processing helix chain 'C' and resid 282 through 287 removed outlier: 3.610A pdb=" N TYR C 286 " --> pdb=" O SER C 282 " (cutoff:3.500A) Processing helix chain 'C' and resid 288 through 310 removed outlier: 3.522A pdb=" N LEU C 292 " --> pdb=" O PHE C 288 " (cutoff:3.500A) Proline residue: C 295 - end of helix Processing helix chain 'D' and resid 72 through 98 Processing helix chain 'D' and resid 103 through 122 removed outlier: 3.678A pdb=" N VAL D 120 " --> pdb=" O MET D 116 " (cutoff:3.500A) Processing helix chain 'D' and resid 130 through 140 removed outlier: 3.508A pdb=" N THR D 134 " --> pdb=" O ASN D 130 " (cutoff:3.500A) removed outlier: 3.703A pdb=" N ALA D 138 " --> pdb=" O THR D 134 " (cutoff:3.500A) removed outlier: 3.589A pdb=" N VAL D 139 " --> pdb=" O VAL D 135 " (cutoff:3.500A) removed outlier: 4.128A pdb=" N PHE D 140 " --> pdb=" O GLY D 136 " (cutoff:3.500A) Processing helix chain 'D' and resid 147 through 170 Processing helix chain 'D' and resid 172 through 177 Processing helix chain 'D' and resid 190 through 195 removed outlier: 4.104A pdb=" N PHE D 194 " --> pdb=" O ILE D 190 " (cutoff:3.500A) Processing helix chain 'D' and resid 204 through 226 removed outlier: 3.659A pdb=" N ILE D 208 " --> pdb=" O ILE D 204 " (cutoff:3.500A) removed outlier: 3.582A pdb=" N THR D 215 " --> pdb=" O GLU D 211 " (cutoff:3.500A) Processing helix chain 'D' and resid 236 through 253 Processing helix chain 'D' and resid 261 through 276 Proline residue: D 268 - end of helix Processing helix chain 'D' and resid 282 through 287 removed outlier: 3.610A pdb=" N TYR D 286 " --> pdb=" O SER D 282 " (cutoff:3.500A) Processing helix chain 'D' and resid 288 through 310 removed outlier: 3.522A pdb=" N LEU D 292 " --> pdb=" O PHE D 288 " (cutoff:3.500A) Proline residue: D 295 - end of helix 481 hydrogen bonds defined for protein. 1443 hydrogen bond angles defined for protein. Restraints generated for nucleic acids: 0 hydrogen bonds 0 hydrogen bond angles 0 basepair planarities 0 basepair parallelities 0 stacking parallelities Total time for adding SS restraints: 1.94 Time building geometry restraints manager: 3.21 seconds NOTE: a complete listing of the restraints can be obtained by requesting output of .geo file. Histogram of bond lengths: 1.23 - 1.34: 2196 1.34 - 1.46: 1735 1.46 - 1.57: 3641 1.57 - 1.69: 0 1.69 - 1.81: 52 Bond restraints: 7624 Sorted by residual: bond pdb=" C9 PNT A 401 " pdb=" N1 PNT A 401 " ideal model delta sigma weight residual 1.279 1.389 -0.110 2.00e-02 2.50e+03 3.03e+01 bond pdb=" C9 PNT B 401 " pdb=" N1 PNT B 401 " ideal model delta sigma weight residual 1.279 1.389 -0.110 2.00e-02 2.50e+03 3.03e+01 bond pdb=" C9 PNT C 401 " pdb=" N1 PNT C 401 " ideal model delta sigma weight residual 1.279 1.389 -0.110 2.00e-02 2.50e+03 3.03e+01 bond pdb=" C9 PNT D 401 " pdb=" N1 PNT D 401 " ideal model delta sigma weight residual 1.279 1.389 -0.110 2.00e-02 2.50e+03 3.01e+01 bond pdb=" C9 PNT A 401 " pdb=" N2 PNT A 401 " ideal model delta sigma weight residual 1.368 1.311 0.057 2.00e-02 2.50e+03 8.23e+00 ... (remaining 7619 not shown) Histogram of bond angle deviations from ideal: 100.14 - 106.90: 236 106.90 - 113.67: 3923 113.67 - 120.44: 3156 120.44 - 127.20: 2997 127.20 - 133.97: 84 Bond angle restraints: 10396 Sorted by residual: angle pdb=" C4 PNT C 401 " pdb=" C9 PNT C 401 " pdb=" N2 PNT C 401 " ideal model delta sigma weight residual 115.38 127.20 -11.82 3.00e+00 1.11e-01 1.55e+01 angle pdb=" C4 PNT B 401 " pdb=" C9 PNT B 401 " pdb=" N2 PNT B 401 " ideal model delta sigma weight residual 115.38 127.20 -11.82 3.00e+00 1.11e-01 1.55e+01 angle pdb=" C4 PNT A 401 " pdb=" C9 PNT A 401 " pdb=" N2 PNT A 401 " ideal model delta sigma weight residual 115.38 127.20 -11.82 3.00e+00 1.11e-01 1.55e+01 angle pdb=" C4 PNT D 401 " pdb=" C9 PNT D 401 " pdb=" N2 PNT D 401 " ideal model delta sigma weight residual 115.38 127.17 -11.79 3.00e+00 1.11e-01 1.55e+01 angle pdb=" N1 PNT D 401 " pdb=" C9 PNT D 401 " pdb=" N2 PNT D 401 " ideal model delta sigma weight residual 125.71 114.42 11.29 3.00e+00 1.11e-01 1.42e+01 ... (remaining 10391 not shown) Histogram of dihedral angle deviations from ideal: 0.00 - 32.36: 4048 32.36 - 64.73: 152 64.73 - 97.09: 12 97.09 - 129.45: 0 129.45 - 161.81: 4 Dihedral angle restraints: 4216 sinusoidal: 1520 harmonic: 2696 Sorted by residual: dihedral pdb=" C10 PNT D 401 " pdb=" C7 PNT D 401 " pdb=" C8 PNT D 401 " pdb=" O1 PNT D 401 " ideal model delta sinusoidal sigma weight residual 180.84 19.03 161.81 1 3.00e+01 1.11e-03 2.08e+01 dihedral pdb=" C10 PNT B 401 " pdb=" C7 PNT B 401 " pdb=" C8 PNT B 401 " pdb=" O1 PNT B 401 " ideal model delta sinusoidal sigma weight residual 180.84 19.04 161.80 1 3.00e+01 1.11e-03 2.08e+01 dihedral pdb=" C10 PNT C 401 " pdb=" C7 PNT C 401 " pdb=" C8 PNT C 401 " pdb=" O1 PNT C 401 " ideal model delta sinusoidal sigma weight residual 180.84 19.06 161.78 1 3.00e+01 1.11e-03 2.08e+01 ... (remaining 4213 not shown) Histogram of chiral volume deviations from ideal: 0.000 - 0.032: 719 0.032 - 0.065: 329 0.065 - 0.097: 74 0.097 - 0.129: 22 0.129 - 0.162: 4 Chirality restraints: 1148 Sorted by residual: chirality pdb=" CA ASN C 137 " pdb=" N ASN C 137 " pdb=" C ASN C 137 " pdb=" CB ASN C 137 " both_signs ideal model delta sigma weight residual False 2.51 2.35 0.16 2.00e-01 2.50e+01 6.54e-01 chirality pdb=" CA ASN D 137 " pdb=" N ASN D 137 " pdb=" C ASN D 137 " pdb=" CB ASN D 137 " both_signs ideal model delta sigma weight residual False 2.51 2.35 0.16 2.00e-01 2.50e+01 6.54e-01 chirality pdb=" CA ASN B 137 " pdb=" N ASN B 137 " pdb=" C ASN B 137 " pdb=" CB ASN B 137 " both_signs ideal model delta sigma weight residual False 2.51 2.35 0.16 2.00e-01 2.50e+01 6.49e-01 ... (remaining 1145 not shown) Planarity restraints: 1308 Sorted by residual: delta sigma weight rms_deltas residual plane pdb=" CA ASN D 137 " -0.013 2.00e-02 2.50e+03 2.57e-02 6.61e+00 pdb=" C ASN D 137 " 0.044 2.00e-02 2.50e+03 pdb=" O ASN D 137 " -0.017 2.00e-02 2.50e+03 pdb=" N ALA D 138 " -0.015 2.00e-02 2.50e+03 delta sigma weight rms_deltas residual plane pdb=" CA ASN C 137 " 0.013 2.00e-02 2.50e+03 2.57e-02 6.61e+00 pdb=" C ASN C 137 " -0.044 2.00e-02 2.50e+03 pdb=" O ASN C 137 " 0.017 2.00e-02 2.50e+03 pdb=" N ALA C 138 " 0.015 2.00e-02 2.50e+03 delta sigma weight rms_deltas residual plane pdb=" CA ASN A 137 " -0.013 2.00e-02 2.50e+03 2.57e-02 6.59e+00 pdb=" C ASN A 137 " 0.044 2.00e-02 2.50e+03 pdb=" O ASN A 137 " -0.017 2.00e-02 2.50e+03 pdb=" N ALA A 138 " -0.015 2.00e-02 2.50e+03 ... (remaining 1305 not shown) Histogram of nonbonded interaction distances: 2.34 - 2.85: 2717 2.85 - 3.37: 7150 3.37 - 3.88: 13091 3.88 - 4.39: 14895 4.39 - 4.90: 25485 Nonbonded interactions: 63338 Sorted by model distance: nonbonded pdb=" O MET B 156 " pdb=" OG1 THR B 159 " model vdw 2.343 2.440 nonbonded pdb=" O MET C 156 " pdb=" OG1 THR C 159 " model vdw 2.344 2.440 nonbonded pdb=" O MET D 156 " pdb=" OG1 THR D 159 " model vdw 2.344 2.440 nonbonded pdb=" O MET A 156 " pdb=" OG1 THR A 159 " model vdw 2.344 2.440 nonbonded pdb=" N ASP A 73 " pdb=" OD1 ASP A 73 " model vdw 2.367 2.520 ... (remaining 63333 not shown) NOTE: a complete listing of the restraints can be obtained by requesting output of .geo file. Find NCS groups from input model Found NCS groups: ncs_group { reference = chain 'A' selection = chain 'B' selection = chain 'C' selection = chain 'D' } Set up NCS constraints No NCS constraints will be used in refinement. Set refine NCS operators Adjust number of macro_cycles Number of macro_cycles: 10 Reset NCS operators Extract rigid body selections Check and reset occupancies Occupancies: min=1.00 max=1.00 mean=1.00 Load rotamer database and sin/cos tables Set ADP refinement strategy ADPs will be refined as individual isotropic Make a string to write initial .geo file Internal consistency checks Time: Set random seed: 0.000 Set model cs if undefined: 0.000 Decide on map wrapping: 0.000 Normalize map: mean=0, sd=1: 0.710 Set stop_for_unknowns flag: 0.000 Assert model is a single copy model: 0.000 Assert all atoms have isotropic ADPs: 0.010 Construct map_model_manager: 0.000 Extract box with map and model: 0.280 Check model and map are aligned: 0.050 Set scattering table: 0.100 Process input model: 23.490 Find NCS groups from input model: 0.380 Set up NCS constraints: 0.030 Set refine NCS operators: 0.000 Adjust number of macro_cycles: 0.000 Reset NCS operators: 0.000 Extract rigid body selections: 0.000 Check and reset occupancies: 0.000 Load rotamer database and sin/cos tables:12.600 Set ADP refinement strategy: 0.000 Make a string to write initial .geo file:0.000 Internal consistency checks: 0.000 Total: 37.650 ------------------------------------------------------------------------------- Set refinement monitor ********************** ------------------------------------------------------------------------------- Setup refinement engine *********************** ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7376 moved from start: 0.0000 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.005 0.110 7624 Z= 0.322 Angle : 0.690 11.821 10396 Z= 0.340 Chirality : 0.038 0.162 1148 Planarity : 0.006 0.030 1308 Dihedral : 17.048 161.815 2536 Min Nonbonded Distance : 2.343 Molprobity Statistics. All-atom Clashscore : 7.64 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.08 % Favored : 97.92 % Rotamer: Outliers : 1.65 % Allowed : 21.43 % Favored : 76.92 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 0.01 (0.27), residues: 960 helix: 0.82 (0.21), residues: 628 sheet: None (None), residues: 0 loop : -1.55 (0.30), residues: 332 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.005 0.001 TRP C 289 HIS 0.001 0.000 HIS C 283 PHE 0.010 0.001 PHE A 246 TYR 0.007 0.001 TYR D 206 ARG 0.002 0.001 ARG D 146 *********************** REFINEMENT MACRO_CYCLE 1 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 146 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 12 poor density : 134 time to evaluate : 0.811 Fit side-chains REVERT: A 172 ASP cc_start: 0.6769 (t0) cc_final: 0.6429 (t0) REVERT: A 196 MET cc_start: 0.7386 (mmt) cc_final: 0.7173 (mmt) REVERT: A 308 TYR cc_start: 0.6031 (t80) cc_final: 0.5784 (t80) REVERT: B 172 ASP cc_start: 0.6718 (t0) cc_final: 0.6394 (t0) REVERT: B 196 MET cc_start: 0.7376 (mmt) cc_final: 0.7165 (mmt) REVERT: B 308 TYR cc_start: 0.6007 (t80) cc_final: 0.5779 (t80) REVERT: C 172 ASP cc_start: 0.6776 (t0) cc_final: 0.6437 (t0) REVERT: C 196 MET cc_start: 0.7430 (mmt) cc_final: 0.7203 (mmt) REVERT: C 308 TYR cc_start: 0.5999 (t80) cc_final: 0.5767 (t80) REVERT: D 172 ASP cc_start: 0.6768 (t0) cc_final: 0.6429 (t0) REVERT: D 196 MET cc_start: 0.7435 (mmt) cc_final: 0.7209 (mmt) REVERT: D 308 TYR cc_start: 0.6035 (t80) cc_final: 0.5781 (t80) outliers start: 12 outliers final: 4 residues processed: 142 average time/residue: 0.8684 time to fit residues: 132.7453 Evaluate side-chains 102 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 4 poor density : 98 time to evaluate : 0.818 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 222 VAL Chi-restraints excluded: chain B residue 222 VAL Chi-restraints excluded: chain C residue 222 VAL Chi-restraints excluded: chain D residue 222 VAL Rotamers are restrained with sigma=5.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 81 optimal weight: 2.9990 chunk 72 optimal weight: 0.9980 chunk 40 optimal weight: 0.9990 chunk 24 optimal weight: 0.9990 chunk 49 optimal weight: 0.7980 chunk 38 optimal weight: 0.9990 chunk 75 optimal weight: 0.5980 chunk 29 optimal weight: 0.6980 chunk 45 optimal weight: 2.9990 chunk 56 optimal weight: 2.9990 chunk 87 optimal weight: 0.8980 overall best weight: 0.7980 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7534 moved from start: 0.1655 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.004 0.049 7624 Z= 0.232 Angle : 0.592 7.918 10396 Z= 0.304 Chirality : 0.039 0.128 1148 Planarity : 0.005 0.037 1308 Dihedral : 12.863 149.767 1080 Min Nonbonded Distance : 2.392 Molprobity Statistics. All-atom Clashscore : 8.25 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.71 % Favored : 97.29 % Rotamer: Outliers : 2.88 % Allowed : 18.82 % Favored : 78.30 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 0.98 (0.27), residues: 960 helix: 1.64 (0.21), residues: 652 sheet: None (None), residues: 0 loop : -1.57 (0.30), residues: 308 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.003 0.001 TRP A 192 HIS 0.002 0.001 HIS B 128 PHE 0.017 0.002 PHE B 246 TYR 0.009 0.001 TYR D 71 ARG 0.002 0.000 ARG C 269 *********************** REFINEMENT MACRO_CYCLE 2 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 144 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 21 poor density : 123 time to evaluate : 0.808 Fit side-chains REVERT: A 145 TRP cc_start: 0.7796 (m100) cc_final: 0.7321 (m100) REVERT: A 258 LEU cc_start: 0.7535 (tp) cc_final: 0.7287 (tp) REVERT: B 145 TRP cc_start: 0.7801 (m100) cc_final: 0.7322 (m100) REVERT: B 258 LEU cc_start: 0.7546 (tp) cc_final: 0.7298 (tp) REVERT: C 145 TRP cc_start: 0.7794 (m100) cc_final: 0.7333 (m100) REVERT: C 258 LEU cc_start: 0.7539 (tp) cc_final: 0.7294 (tp) REVERT: D 145 TRP cc_start: 0.7801 (m100) cc_final: 0.7337 (m100) REVERT: D 258 LEU cc_start: 0.7535 (tp) cc_final: 0.7281 (tp) outliers start: 21 outliers final: 7 residues processed: 131 average time/residue: 0.8860 time to fit residues: 124.6746 Evaluate side-chains 119 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 7 poor density : 112 time to evaluate : 0.813 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 222 VAL Chi-restraints excluded: chain B residue 183 ILE Chi-restraints excluded: chain B residue 222 VAL Chi-restraints excluded: chain C residue 222 VAL Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 222 VAL Rotamers are restrained with sigma=4.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 48 optimal weight: 2.9990 chunk 27 optimal weight: 1.9990 chunk 72 optimal weight: 2.9990 chunk 59 optimal weight: 1.9990 chunk 24 optimal weight: 2.9990 chunk 87 optimal weight: 0.4980 chunk 94 optimal weight: 1.9990 chunk 77 optimal weight: 2.9990 chunk 86 optimal weight: 0.9990 chunk 29 optimal weight: 1.9990 chunk 70 optimal weight: 1.9990 overall best weight: 1.4988 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 250 ASN B 250 ASN C 250 ASN D 250 ASN Total number of N/Q/H flips: 4 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7605 moved from start: 0.2602 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.005 0.054 7624 Z= 0.335 Angle : 0.676 8.775 10396 Z= 0.350 Chirality : 0.041 0.119 1148 Planarity : 0.006 0.036 1308 Dihedral : 13.376 159.949 1080 Min Nonbonded Distance : 2.429 Molprobity Statistics. All-atom Clashscore : 7.03 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.92 % Favored : 97.08 % Rotamer: Outliers : 5.77 % Allowed : 13.05 % Favored : 81.18 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 0.61 (0.26), residues: 960 helix: 1.28 (0.20), residues: 656 sheet: None (None), residues: 0 loop : -1.56 (0.32), residues: 304 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.003 0.001 TRP A 192 HIS 0.004 0.001 HIS A 177 PHE 0.020 0.003 PHE A 246 TYR 0.008 0.002 TYR D 197 ARG 0.001 0.000 ARG D 72 *********************** REFINEMENT MACRO_CYCLE 3 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 160 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 42 poor density : 118 time to evaluate : 0.830 Fit side-chains REVERT: C 279 GLU cc_start: 0.7559 (OUTLIER) cc_final: 0.7256 (pm20) REVERT: D 188 LYS cc_start: 0.6740 (ptpp) cc_final: 0.6447 (ptpt) outliers start: 42 outliers final: 17 residues processed: 138 average time/residue: 1.0737 time to fit residues: 157.7226 Evaluate side-chains 130 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 18 poor density : 112 time to evaluate : 0.770 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 103 LEU Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain A residue 222 VAL Chi-restraints excluded: chain B residue 103 LEU Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain B residue 222 VAL Chi-restraints excluded: chain C residue 103 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 222 VAL Chi-restraints excluded: chain C residue 279 GLU Chi-restraints excluded: chain D residue 103 LEU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Chi-restraints excluded: chain D residue 222 VAL Rotamers are restrained with sigma=4.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 86 optimal weight: 0.8980 chunk 65 optimal weight: 1.9990 chunk 45 optimal weight: 0.0470 chunk 9 optimal weight: 4.9990 chunk 41 optimal weight: 1.9990 chunk 58 optimal weight: 2.9990 chunk 87 optimal weight: 0.3980 chunk 92 optimal weight: 0.0010 chunk 83 optimal weight: 5.9990 chunk 25 optimal weight: 1.9990 chunk 77 optimal weight: 0.0470 overall best weight: 0.2782 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7522 moved from start: 0.2464 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.003 0.050 7624 Z= 0.166 Angle : 0.535 8.935 10396 Z= 0.266 Chirality : 0.037 0.112 1148 Planarity : 0.005 0.037 1308 Dihedral : 13.153 165.663 1080 Min Nonbonded Distance : 2.378 Molprobity Statistics. All-atom Clashscore : 7.98 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.60 % Favored : 97.40 % Rotamer: Outliers : 2.06 % Allowed : 16.90 % Favored : 81.04 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.42 (0.27), residues: 960 helix: 2.02 (0.20), residues: 636 sheet: None (None), residues: 0 loop : -1.33 (0.31), residues: 324 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.005 0.001 TRP A 289 HIS 0.001 0.000 HIS D 177 PHE 0.010 0.001 PHE A 246 TYR 0.006 0.001 TYR D 236 ARG 0.002 0.000 ARG B 269 *********************** REFINEMENT MACRO_CYCLE 4 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 128 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 15 poor density : 113 time to evaluate : 0.772 Fit side-chains REVERT: A 222 VAL cc_start: 0.7439 (t) cc_final: 0.7203 (t) REVERT: B 222 VAL cc_start: 0.7433 (t) cc_final: 0.7199 (t) REVERT: C 222 VAL cc_start: 0.7441 (t) cc_final: 0.7208 (t) REVERT: C 279 GLU cc_start: 0.7481 (OUTLIER) cc_final: 0.7245 (pm20) REVERT: D 222 VAL cc_start: 0.7430 (t) cc_final: 0.7197 (t) outliers start: 15 outliers final: 9 residues processed: 122 average time/residue: 0.9348 time to fit residues: 122.2650 Evaluate side-chains 115 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 10 poor density : 105 time to evaluate : 0.764 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 279 GLU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Rotamers are restrained with sigma=3.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 52 optimal weight: 0.9990 chunk 1 optimal weight: 0.8980 chunk 69 optimal weight: 0.7980 chunk 38 optimal weight: 0.7980 chunk 79 optimal weight: 3.9990 chunk 64 optimal weight: 0.6980 chunk 0 optimal weight: 3.9990 chunk 47 optimal weight: 0.5980 chunk 83 optimal weight: 0.0070 chunk 23 optimal weight: 2.9990 chunk 31 optimal weight: 2.9990 overall best weight: 0.5798 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7549 moved from start: 0.2557 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.003 0.051 7624 Z= 0.195 Angle : 0.558 8.513 10396 Z= 0.280 Chirality : 0.037 0.115 1148 Planarity : 0.005 0.038 1308 Dihedral : 12.969 164.173 1076 Min Nonbonded Distance : 2.417 Molprobity Statistics. All-atom Clashscore : 7.58 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.92 % Favored : 97.08 % Rotamer: Outliers : 3.16 % Allowed : 17.03 % Favored : 79.81 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.50 (0.27), residues: 960 helix: 2.09 (0.20), residues: 636 sheet: None (None), residues: 0 loop : -1.32 (0.31), residues: 324 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.003 0.001 TRP A 289 HIS 0.002 0.001 HIS A 177 PHE 0.016 0.002 PHE D 246 TYR 0.006 0.001 TYR B 236 ARG 0.001 0.000 ARG A 269 *********************** REFINEMENT MACRO_CYCLE 5 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 134 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 23 poor density : 111 time to evaluate : 0.768 Fit side-chains REVERT: C 258 LEU cc_start: 0.7947 (OUTLIER) cc_final: 0.7465 (mt) REVERT: C 279 GLU cc_start: 0.7522 (OUTLIER) cc_final: 0.7259 (pm20) REVERT: D 258 LEU cc_start: 0.7943 (OUTLIER) cc_final: 0.7530 (mt) outliers start: 23 outliers final: 11 residues processed: 125 average time/residue: 0.9585 time to fit residues: 128.2576 Evaluate side-chains 125 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 14 poor density : 111 time to evaluate : 0.716 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 96 LEU Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain B residue 146 ARG Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 258 LEU Chi-restraints excluded: chain C residue 279 GLU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Chi-restraints excluded: chain D residue 258 LEU Rotamers are restrained with sigma=3.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 83 optimal weight: 0.0770 chunk 18 optimal weight: 0.0970 chunk 54 optimal weight: 1.9990 chunk 22 optimal weight: 0.0030 chunk 92 optimal weight: 0.6980 chunk 77 optimal weight: 5.9990 chunk 43 optimal weight: 2.9990 chunk 7 optimal weight: 0.4980 chunk 30 optimal weight: 0.6980 chunk 48 optimal weight: 1.9990 chunk 89 optimal weight: 0.9980 overall best weight: 0.2746 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7501 moved from start: 0.2507 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.002 0.050 7624 Z= 0.161 Angle : 0.526 7.978 10396 Z= 0.259 Chirality : 0.036 0.111 1148 Planarity : 0.005 0.039 1308 Dihedral : 12.699 163.490 1076 Min Nonbonded Distance : 2.390 Molprobity Statistics. All-atom Clashscore : 7.85 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.60 % Favored : 97.40 % Rotamer: Outliers : 3.02 % Allowed : 17.17 % Favored : 79.81 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.78 (0.27), residues: 960 helix: 2.37 (0.20), residues: 636 sheet: None (None), residues: 0 loop : -1.34 (0.30), residues: 324 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.005 0.001 TRP A 289 HIS 0.001 0.000 HIS A 177 PHE 0.008 0.001 PHE D 246 TYR 0.007 0.001 TYR A 236 ARG 0.001 0.000 ARG B 269 *********************** REFINEMENT MACRO_CYCLE 6 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 130 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 22 poor density : 108 time to evaluate : 0.793 Fit side-chains REVERT: A 222 VAL cc_start: 0.7432 (t) cc_final: 0.7137 (t) REVERT: B 222 VAL cc_start: 0.7423 (t) cc_final: 0.7133 (t) REVERT: C 222 VAL cc_start: 0.7433 (t) cc_final: 0.7140 (t) REVERT: C 279 GLU cc_start: 0.7477 (OUTLIER) cc_final: 0.7218 (pm20) REVERT: D 222 VAL cc_start: 0.7422 (t) cc_final: 0.7141 (t) outliers start: 22 outliers final: 13 residues processed: 123 average time/residue: 0.9899 time to fit residues: 130.1899 Evaluate side-chains 117 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 14 poor density : 103 time to evaluate : 0.780 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 73 ASP Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain B residue 73 ASP Chi-restraints excluded: chain B residue 146 ARG Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 279 GLU Chi-restraints excluded: chain D residue 73 ASP Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Rotamers are restrained with sigma=2.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 10 optimal weight: 0.6980 chunk 52 optimal weight: 1.9990 chunk 67 optimal weight: 3.9990 chunk 78 optimal weight: 1.9990 chunk 51 optimal weight: 0.5980 chunk 92 optimal weight: 4.9990 chunk 57 optimal weight: 1.9990 chunk 56 optimal weight: 0.9980 chunk 42 optimal weight: 0.0970 chunk 36 optimal weight: 0.8980 chunk 55 optimal weight: 2.9990 overall best weight: 0.6578 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7550 moved from start: 0.2627 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.003 0.050 7624 Z= 0.202 Angle : 0.567 8.272 10396 Z= 0.283 Chirality : 0.037 0.116 1148 Planarity : 0.005 0.040 1308 Dihedral : 12.867 165.446 1076 Min Nonbonded Distance : 2.408 Molprobity Statistics. All-atom Clashscore : 8.39 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.92 % Favored : 97.08 % Rotamer: Outliers : 3.43 % Allowed : 15.93 % Favored : 80.63 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.68 (0.27), residues: 960 helix: 2.23 (0.20), residues: 640 sheet: None (None), residues: 0 loop : -1.29 (0.31), residues: 320 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.002 0.001 TRP A 289 HIS 0.002 0.001 HIS A 177 PHE 0.016 0.002 PHE D 246 TYR 0.007 0.001 TYR A 236 ARG 0.001 0.000 ARG A 72 *********************** REFINEMENT MACRO_CYCLE 7 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 131 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 25 poor density : 106 time to evaluate : 0.781 Fit side-chains REVERT: A 188 LYS cc_start: 0.6863 (ptpp) cc_final: 0.6631 (ptpt) REVERT: C 279 GLU cc_start: 0.7520 (OUTLIER) cc_final: 0.7253 (pm20) outliers start: 25 outliers final: 10 residues processed: 120 average time/residue: 0.9709 time to fit residues: 124.5167 Evaluate side-chains 117 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 11 poor density : 106 time to evaluate : 0.767 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain B residue 146 ARG Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 279 GLU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Rotamers are restrained with sigma=2.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 27 optimal weight: 0.7980 chunk 18 optimal weight: 2.9990 chunk 17 optimal weight: 0.9980 chunk 58 optimal weight: 2.9990 chunk 63 optimal weight: 0.9990 chunk 45 optimal weight: 2.9990 chunk 8 optimal weight: 0.5980 chunk 72 optimal weight: 0.6980 chunk 84 optimal weight: 1.9990 chunk 88 optimal weight: 0.6980 chunk 80 optimal weight: 1.9990 overall best weight: 0.7580 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7562 moved from start: 0.2763 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.003 0.051 7624 Z= 0.215 Angle : 0.595 8.536 10396 Z= 0.295 Chirality : 0.038 0.116 1148 Planarity : 0.005 0.039 1308 Dihedral : 12.881 166.068 1076 Min Nonbonded Distance : 2.407 Molprobity Statistics. All-atom Clashscore : 8.12 Ramachandran Plot: Outliers : 0.00 % Allowed : 3.02 % Favored : 96.98 % Rotamer: Outliers : 3.71 % Allowed : 15.66 % Favored : 80.63 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.60 (0.26), residues: 960 helix: 2.24 (0.20), residues: 632 sheet: None (None), residues: 0 loop : -1.38 (0.31), residues: 328 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.002 0.001 TRP C 192 HIS 0.003 0.001 HIS A 177 PHE 0.017 0.002 PHE A 246 TYR 0.008 0.001 TYR B 206 ARG 0.001 0.000 ARG A 72 *********************** REFINEMENT MACRO_CYCLE 8 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 132 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 27 poor density : 105 time to evaluate : 0.841 Fit side-chains REVERT: A 146 ARG cc_start: 0.7548 (OUTLIER) cc_final: 0.7222 (tmm160) REVERT: B 258 LEU cc_start: 0.8048 (OUTLIER) cc_final: 0.7574 (mt) REVERT: C 258 LEU cc_start: 0.8040 (OUTLIER) cc_final: 0.7565 (mt) outliers start: 27 outliers final: 11 residues processed: 119 average time/residue: 0.9568 time to fit residues: 121.9289 Evaluate side-chains 117 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 14 poor density : 103 time to evaluate : 0.760 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 146 ARG Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain B residue 96 LEU Chi-restraints excluded: chain B residue 146 ARG Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain B residue 258 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 258 LEU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Rotamers are restrained with sigma=1.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 86 optimal weight: 0.9980 chunk 88 optimal weight: 2.9990 chunk 51 optimal weight: 2.9990 chunk 37 optimal weight: 4.9990 chunk 67 optimal weight: 2.9990 chunk 26 optimal weight: 0.7980 chunk 77 optimal weight: 0.5980 chunk 81 optimal weight: 1.9990 chunk 85 optimal weight: 0.9980 chunk 56 optimal weight: 1.9990 chunk 91 optimal weight: 1.9990 overall best weight: 1.0782 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7588 moved from start: 0.2919 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.004 0.051 7624 Z= 0.264 Angle : 0.635 8.464 10396 Z= 0.319 Chirality : 0.040 0.116 1148 Planarity : 0.006 0.039 1308 Dihedral : 12.957 167.072 1076 Min Nonbonded Distance : 2.407 Molprobity Statistics. All-atom Clashscore : 6.97 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.92 % Favored : 97.08 % Rotamer: Outliers : 2.47 % Allowed : 16.76 % Favored : 80.77 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.48 (0.27), residues: 960 helix: 2.10 (0.19), residues: 632 sheet: None (None), residues: 0 loop : -1.33 (0.33), residues: 328 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.002 0.001 TRP A 192 HIS 0.003 0.001 HIS A 177 PHE 0.017 0.002 PHE A 246 TYR 0.009 0.001 TYR C 206 ARG 0.001 0.000 ARG A 72 *********************** REFINEMENT MACRO_CYCLE 9 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 132 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 18 poor density : 114 time to evaluate : 0.863 Fit side-chains REVERT: A 146 ARG cc_start: 0.7530 (OUTLIER) cc_final: 0.7195 (tmm160) REVERT: A 188 LYS cc_start: 0.6946 (ptpt) cc_final: 0.6621 (ptpt) REVERT: A 258 LEU cc_start: 0.7998 (OUTLIER) cc_final: 0.7733 (mt) REVERT: C 258 LEU cc_start: 0.8045 (OUTLIER) cc_final: 0.7677 (mt) REVERT: D 188 LYS cc_start: 0.6766 (ptpp) cc_final: 0.6508 (ptpt) outliers start: 18 outliers final: 10 residues processed: 123 average time/residue: 1.0455 time to fit residues: 136.7605 Evaluate side-chains 124 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 13 poor density : 111 time to evaluate : 0.789 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 146 ARG Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain A residue 258 LEU Chi-restraints excluded: chain B residue 146 ARG Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 258 LEU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Rotamers are restrained with sigma=1.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 55 optimal weight: 0.0470 chunk 43 optimal weight: 1.9990 chunk 63 optimal weight: 0.9990 chunk 95 optimal weight: 0.6980 chunk 88 optimal weight: 3.9990 chunk 76 optimal weight: 1.9990 chunk 7 optimal weight: 0.9980 chunk 58 optimal weight: 2.9990 chunk 46 optimal weight: 0.8980 chunk 60 optimal weight: 0.6980 chunk 81 optimal weight: 1.9990 overall best weight: 0.6678 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7557 moved from start: 0.2858 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.003 0.050 7624 Z= 0.204 Angle : 0.584 8.645 10396 Z= 0.289 Chirality : 0.038 0.115 1148 Planarity : 0.005 0.039 1308 Dihedral : 12.786 166.808 1076 Min Nonbonded Distance : 2.392 Molprobity Statistics. All-atom Clashscore : 8.12 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.50 % Favored : 97.50 % Rotamer: Outliers : 2.75 % Allowed : 16.90 % Favored : 80.36 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.70 (0.26), residues: 960 helix: 2.32 (0.19), residues: 632 sheet: None (None), residues: 0 loop : -1.38 (0.32), residues: 328 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.003 0.001 TRP A 289 HIS 0.002 0.001 HIS A 177 PHE 0.015 0.002 PHE A 246 TYR 0.008 0.001 TYR D 236 ARG 0.001 0.000 ARG A 269 ********************** REFINEMENT MACRO_CYCLE 10 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 1920 Ramachandran restraints generated. 960 Oldfield, 0 Emsley, 960 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 124 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 20 poor density : 104 time to evaluate : 0.713 Fit side-chains REVERT: A 146 ARG cc_start: 0.7539 (OUTLIER) cc_final: 0.7202 (tmm160) REVERT: C 258 LEU cc_start: 0.8020 (OUTLIER) cc_final: 0.7534 (mt) REVERT: D 188 LYS cc_start: 0.6746 (ptpp) cc_final: 0.6495 (ptpt) outliers start: 20 outliers final: 10 residues processed: 116 average time/residue: 0.9874 time to fit residues: 122.4415 Evaluate side-chains 116 residues out of total 728 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 12 poor density : 104 time to evaluate : 0.713 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Chi-restraints excluded: chain A residue 146 ARG Chi-restraints excluded: chain A residue 157 LEU Chi-restraints excluded: chain A residue 219 LEU Chi-restraints excluded: chain B residue 146 ARG Chi-restraints excluded: chain B residue 157 LEU Chi-restraints excluded: chain B residue 219 LEU Chi-restraints excluded: chain C residue 157 LEU Chi-restraints excluded: chain C residue 219 LEU Chi-restraints excluded: chain C residue 258 LEU Chi-restraints excluded: chain D residue 146 ARG Chi-restraints excluded: chain D residue 157 LEU Chi-restraints excluded: chain D residue 219 LEU Rotamers are restrained with sigma=1.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 96 random chunks: chunk 23 optimal weight: 2.9990 chunk 70 optimal weight: 0.7980 chunk 11 optimal weight: 4.9990 chunk 21 optimal weight: 1.9990 chunk 76 optimal weight: 0.9980 chunk 31 optimal weight: 0.0870 chunk 78 optimal weight: 1.9990 chunk 9 optimal weight: 4.9990 chunk 14 optimal weight: 0.5980 chunk 66 optimal weight: 0.8980 chunk 4 optimal weight: 2.9990 overall best weight: 0.6758 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... No N/Q/H corrections needed this macrocycle ------------------------------------------------------------------------------- ADP refinement ************** |-group b-factor refinement (macro cycle = 0; iterations = 0)-----------------| | r_work = 0.3956 r_free = 0.3956 target = 0.181913 restraints weight = None | |-----------------------------------------------------------------------------| |-group b-factor refinement (macro cycle = 1; iterations = 47)----------------| | r_work = 0.3709 r_free = 0.3709 target = 0.158547 restraints weight = 6821.465| |-----------------------------------------------------------------------------| r_work (start): 0.3705 rms_B_bonded: 1.21 r_work: 0.3592 rms_B_bonded: 1.95 restraints_weight: 0.5000 r_work: 0.3487 rms_B_bonded: 3.26 restraints_weight: 0.2500 r_work (final): 0.3487 ------------------------------------------------------------------------------- Occupancy refinement ******************** ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.8010 moved from start: 0.2870 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd, rmsZ for bonds and angles. Bond : 0.003 0.049 7624 Z= 0.204 Angle : 0.587 8.509 10396 Z= 0.291 Chirality : 0.038 0.115 1148 Planarity : 0.005 0.039 1308 Dihedral : 12.777 167.314 1076 Min Nonbonded Distance : 2.398 Molprobity Statistics. All-atom Clashscore : 7.78 Ramachandran Plot: Outliers : 0.00 % Allowed : 2.92 % Favored : 97.08 % Rotamer: Outliers : 1.65 % Allowed : 18.41 % Favored : 79.95 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.00 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: 1.76 (0.26), residues: 960 helix: 2.38 (0.19), residues: 632 sheet: None (None), residues: 0 loop : -1.37 (0.32), residues: 328 Max deviation from planes: Type MaxDev MeanDev LineInFile TRP 0.003 0.001 TRP A 289 HIS 0.002 0.001 HIS A 177 PHE 0.016 0.002 PHE A 246 TYR 0.008 0.001 TYR C 206 ARG 0.001 0.000 ARG A 72 Origin is already at (0, 0, 0), no shifts will be applied =============================================================================== Job complete usr+sys time: 2800.13 seconds wall clock time: 49 minutes 34.91 seconds (2974.91 seconds total)