Full wwPDB X-ray Structure Validation Report i...Total C N O P 28 10 5 11 2 0 0 • Molecule 4 is pyrazinoquinolinone (three-letter code: O7K) (formula: C 25H 23ClF 2N 4O 3) (labeledas"LigandofInterest"byauthor)

Full wwPDB X-ray Structure Validation Report iO

Sep 19, 2020 � 10:03 AM BST

PDB ID : 6T5BTitle : KRasG12C ligand complex

Authors : Phillips, C.Deposited on : 2019-10-15Resolution : 1.37 Å(reported)

This is a Full wwPDB X-ray Structure Validation Report for a publicly released PDB entry.

We welcome your comments at [email protected]

A user guide is available athttps://www.wwpdb.org/validation/2017/XrayValidationReportHelp

with speci�c help available everywhere you see the iO symbol.

The following versions of software and data (see references iO) were used in the production of this report:

MolProbity : 4.02b-467Mogul : 1.8.5 (274361), CSD as541be (2020)

Xtriage (Phenix) : 1.13EDS : 2.14.6

buster-report : 1.1.7 (2018)Percentile statistics : 20191225.v01 (using entries in the PDB archive December 25th 2019)

Refmac : 5.8.0158CCP4 : 7.0.044 (Gargrove)

Ideal geometry (proteins) : Engh & Huber (2001)Ideal geometry (DNA, RNA) : Parkinson et al. (1996)

Validation Pipeline (wwPDB-VP) : 2.14.6

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp



https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#references

Page 2 Full wwPDB X-ray Structure Validation Report 6T5B

1 Overall quality at a glance iO

The following experimental techniques were used to determine the structure:X-RAY DIFFRACTION

The reported resolution of this entry is 1.37 Å.

Percentile scores (ranging between 0-100) for global validation metrics of the entry are shown inthe following graphic. The table shows the number of entries on which the scores are based.

MetricWhole archive(#Entries)

Similar resolution(#Entries, resolution range(Å))

Rfree 130704 2907 (1.40-1.36)Clashscore 141614 3037 (1.40-1.36)

Ramachandran outliers 138981 2970 (1.40-1.36)Sidechain outliers 138945 2969 (1.40-1.36)RSRZ outliers 127900 2846 (1.40-1.36)

The table below summarises the geometric issues observed across the polymeric chains and their�t to the electron density. The red, orange, yellow and green segments on the lower bar indicatethe fraction of residues that contain outliers for >=3, 2, 1 and 0 types of geometric qualitycriteria respectively. A grey segment represents the fraction of residues that are not modelled.The numeric value for each fraction is indicated below the corresponding segment, with a dotrepresenting fractions <=5% The upper red bar (where present) indicates the fraction of residuesthat have poor �t to the electron density. The numeric value is given above the bar.

Mol Chain Length Quality of chain

1 A 170

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#overall_quality


2 Entry composition iO

There are 6 unique types of molecules in this entry. The entry contains 1670 atoms, of which 0are hydrogens and 0 are deuteriums.

In the tables below, the ZeroOcc column contains the number of atoms modelled with zero occu-pancy, the AltConf column contains the number of residues with at least one atom in alternateconformation and the Trace column contains the number of residues modelled with at most 2atoms.

� Molecule 1 is a protein called GTPase KRas.

Mol Chain Residues Atoms ZeroOcc AltConf Trace

1 A 170Total C N O S1359 852 233 269 5

0 0 0

There are 12 discrepancies between the modelled and reference sequences:

Chain Residue Modelled Actual Comment ReferenceA 0 GLY - expression tag UNP P01116A 12 CYS GLY engineered mutation UNP P01116A 51 SER CYS con�ict UNP P01116A 80 LEU CYS con�ict UNP P01116A 118 SER CYS con�ict UNP P01116A 151 GLY ARG con�ict UNP P01116A 153 ASP GLU con�ict UNP P01116A 160 ILE VAL con�ict UNP P01116A 165 LYS GLN con�ict UNP P01116A 166 HIS TYR con�ict UNP P01116A 167 LYS ARG con�ict UNP P01116A 168 GLU LEU con�ict UNP P01116

� Molecule 2 is MAGNESIUM ION (three-letter code: MG) (formula: Mg).

Mol Chain Residues Atoms ZeroOcc AltConf

2 A 1Total Mg1 1

0 0

� Molecule 3 is GUANOSINE-5'-DIPHOSPHATE (three-letter code: GDP) (formula:C10H15N5O11P2).

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#entry_composition



3 A 1Total C N O P28 10 5 11 2

0 0

� Molecule 4 is pyrazinoquinolinone (three-letter code: O7K) (formula: C25H23ClF2N4O3)(labeled as "Ligand of Interest" by author).


4 A 1Total C Cl F N O35 25 1 2 4 3

0 0

� Molecule 5 is SULFATE ION (three-letter code: SO4) (formula: O4S).



5 A 1Total O S5 4 1

0 0

� Molecule 6 is water.


6 A 242Total O242 242

0 0


3 Residue-property plots iO

These plots are drawn for all protein, RNA, DNA and oligosaccharide chains in the entry. The�rst graphic for a chain summarises the proportions of the various outlier classes displayed in thesecond graphic. The second graphic shows the sequence view annotated by issues in geometry andelectron density. Residues are color-coded according to the number of geometric quality criteriafor which they contain at least one outlier: green = 0, yellow = 1, orange = 2 and red = 3 or more.A red dot above a residue indicates a poor �t to the electron density (RSRZ > 2). Stretches of 2or more consecutive residues without any outlier are shown as a green connector. Residues presentin the sample, but not in the model, are shown in grey.

• Molecule 1: GTPase KRas

Chain A:

G0•

M1

T2•

L6

Q25

N26

H27

E31•

E37

R41

L53

D54

I55

L56

A59

G60

Q61

E62

E63

R73

L80

R97

I100

K101

R102

E107•

D108•

R123

D126

T127

K128

D132

R135

R149

F156

I160

K169

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#residue_plots


4 Data and re�nement statistics iO

Property Value SourceSpace group P 1 DepositorCell constantsa, b, c, α, β, γ

32.97Å 36.57Å 37.13Å65.49◦ 74.17◦ 77.61◦

Depositor

Resolution (Å)31.51 � 1.3731.51 � 1.37

DepositorEDS

% Data completeness(in resolution range)

62.2 (31.51-1.37)62.2 (31.51-1.37)

DepositorEDS

Rmerge 0.03 DepositorRsym (Not available) Depositor

< I/σ(I) > 1 49.90 (at 1.37Å) XtriageRe�nement program REFMAC 5.8.0135 Depositor

R, Rfree0.183 , 0.2310.184 , 0.227

DepositorDCC

Rfree test set 964 re�ections (4.90%) wwPDB-VPWilson B-factor (Å2) 7.9 Xtriage

Anisotropy 0.041 XtriageBulk solvent ksol(e/Å3), Bsol(Å2) 0.47 , 59.3 EDS

L-test for twinning2 < |L| > = 0.59, < L2 > = 0.44 XtriageEstimated twinning fraction No twinning to report. Xtriage

Fo,Fc correlation 0.95 EDSTotal number of atoms 1670 wwPDB-VP

Average B, all atoms (Å2) 11.0 wwPDB-VP

Xtriage's analysis on translational NCS is as follows: The analyses of the Patterson function reveals

a signi�cant o�-origin peak that is 21.53 % of the origin peak, indicating pseudo-translational

symmetry. The chance of �nding a peak of this or larger height randomly in a structure without

pseudo-translational symmetry is equal to 6.8815e-03. The detected translational NCS is most

likely also responsible for the elevated intensity ratio.

1Intensities estimated from amplitudes.2Theoretical values of < |L| >, < L2 > for acentric re�ections are 0.5, 0.333 respectively for untwinned datasets,

and 0.375, 0.2 for perfectly twinned datasets.

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#data_stats


5 Model quality iO

5.1 Standard geometry iO

Bond lengths and bond angles in the following residue types are not validated in this section: GDP,MG, SO4, O7K

The Z score for a bond length (or angle) is the number of standard deviations the observed valueis removed from the expected value. A bond length (or angle) with |Z| > 5 is considered anoutlier worth inspection. RMSZ is the root-mean-square of all Z scores of the bond lengths (orangles).

Mol ChainBond lengths Bond anglesRMSZ #|Z| >5 RMSZ #|Z| >5

1 A 1.13 0/1380 1.20 12/1860 (0.6%)

There are no bond length outliers.

All (12) bond angle outliers are listed below:

Mol Chain Res Type Atoms Z Observed(o) Ideal(o)1 A 97 ARG NE-CZ-NH1 -9.68 115.46 120.301 A 126 ASP CB-CG-OD2 -9.40 109.84 118.301 A 123 ARG NE-CZ-NH2 -7.69 116.45 120.301 A 149 ARG NE-CZ-NH1 7.28 123.94 120.301 A 6 LEU CB-CG-CD1 6.58 122.18 111.001 A 135 ARG NE-CZ-NH1 -6.37 117.11 120.301 A 97 ARG NE-CZ-NH2 6.07 123.34 120.301 A 73 ARG NE-CZ-NH2 -6.01 117.29 120.301 A 149 ARG NE-CZ-NH2 -5.88 117.36 120.301 A 126 ASP CB-CG-OD1 5.64 123.37 118.301 A 102 ARG NE-CZ-NH1 5.54 123.07 120.301 A 132 ASP CB-CG-OD1 5.13 122.92 118.30

There are no chirality outliers.

There are no planarity outliers.

5.2 Too-close contacts iO

In the following table, the Non-H and H(model) columns list the number of non-hydrogen atomsand hydrogen atoms in the chain respectively. The H(added) column lists the number of hydrogenatoms added and optimized by MolProbity. The Clashes column lists the number of clashes withinthe asymmetric unit, whereas Symm-Clashes lists symmetry related clashes.

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#model_quality

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#standard_geometry

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#close_contacts


Mol Chain Non-H H(model) H(added) Clashes Symm-Clashes1 A 1359 0 1345 20 02 A 1 0 0 0 03 A 28 0 12 0 04 A 35 0 0 0 05 A 5 0 0 0 06 A 242 0 0 12 0All All 1670 0 1357 20 0

The all-atom clashscore is de�ned as the number of clashes found per 1000 atoms (includinghydrogen atoms). The all-atom clashscore for this structure is 7.

All (20) close contacts within the same asymmetric unit are listed below, sorted by their clashmagnitude.

Atom-1 Atom-2Interatomicdistance (Å)

Clashoverlap (Å)

1:A:56:LEU:HD11 6:A:433:HOH:O 1.61 1.001:A:6:LEU:HD22 6:A:542:HOH:O 1.67 0.951:A:37:GLU:OE2 6:A:301:HOH:O 1.86 0.931:A:37:GLU:CG 6:A:433:HOH:O 2.24 0.841:A:56:LEU:CD1 6:A:433:HOH:O 2.21 0.831:A:156:PHE:HB3 6:A:530:HOH:O 1.87 0.751:A:53:LEU:HD13 6:A:542:HOH:O 1.87 0.751:A:25:GLN:HE21 1:A:27:HIS:HE1 1.35 0.721:A:55:ILE:HD11 6:A:542:HOH:O 1.97 0.631:A:25:GLN:HE21 1:A:27:HIS:CE1 2.22 0.541:A:37:GLU:CD 6:A:433:HOH:O 2.44 0.521:A:6:LEU:HD11 1:A:160:ILE:CD1 2.42 0.501:A:53:LEU:HD22 1:A:160:ILE:HD11 1.95 0.491:A:37:GLU:HG3 6:A:433:HOH:O 2.02 0.481:A:80:LEU:HD21 1:A:100:ILE:CD1 2.44 0.471:A:61:GLN:NE2 1:A:63:GLU:CD 2.69 0.461:A:55:ILE:CD1 6:A:542:HOH:O 2.63 0.431:A:6:LEU:HD21 1:A:160:ILE:HD11 2.00 0.431:A:53:LEU:HB3 6:A:542:HOH:O 2.18 0.421:A:53:LEU:HD22 1:A:160:ILE:CD1 2.50 0.41

There are no symmetry-related clashes.


5.3 Torsion angles iO

5.3.1 Protein backbone iO

In the following table, the Percentiles column shows the percent Ramachandran outliers of thechain as a percentile score with respect to all X-ray entries followed by that with respect to entriesof similar resolution.

The Analysed column shows the number of residues for which the backbone conformation wasanalysed, and the total number of residues.

Mol Chain Analysed Favoured Allowed Outliers Percentiles

1 A 168/170 (99%) 165 (98%) 2 (1%) 1 (1%) 25 7

All (1) Ramachandran outliers are listed below:

Mol Chain Res Type1 A 59 ALA

5.3.2 Protein sidechains iO

In the following table, the Percentiles column shows the percent sidechain outliers of the chain as apercentile score with respect to all X-ray entries followed by that with respect to entries of similarresolution.

The Analysed column shows the number of residues for which the sidechain conformation wasanalysed, and the total number of residues.

Mol Chain Analysed Rotameric Outliers Percentiles

1 A 150/150 (100%) 147 (98%) 3 (2%) 55 23

All (3) residues with a non-rotameric sidechain are listed below:

Mol Chain Res Type1 A 27 HIS1 A 41 ARG1 A 128 LYS

Some sidechains can be �ipped to improve hydrogen bonding and reduce clashes. All (4) suchsidechains are listed below:

Mol Chain Res Type1 A 26 ASN1 A 27 HIS

Continued on next page...

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#torsion_angles

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#protein_backbone

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#protein_sidechains


Continued from previous page...

Mol Chain Res Type1 A 43 GLN1 A 61 GLN

5.3.3 RNA iO

There are no RNA molecules in this entry.

5.4 Non-standard residues in protein, DNA, RNA chains iO

There are no non-standard protein/DNA/RNA residues in this entry.

5.5 Carbohydrates iO

There are no monosaccharides in this entry.

5.6 Ligand geometry iO

Of 4 ligands modelled in this entry, 1 is monoatomic - leaving 3 for Mogul analysis.

In the following table, the Counts columns list the number of bonds (or angles) for which Mogulstatistics could be retrieved, the number of bonds (or angles) that are observed in the model andthe number of bonds (or angles) that are de�ned in the Chemical Component Dictionary. TheLink column lists molecule types, if any, to which the group is linked. The Z score for a bondlength (or angle) is the number of standard deviations the observed value is removed from theexpected value. A bond length (or angle) with |Z| > 2 is considered an outlier worth inspection.RMSZ is the root-mean-square of all Z scores of the bond lengths (or angles).

Mol Type Chain Res LinkBond lengths Bond angles

Counts RMSZ #|Z| > 2 Counts RMSZ #|Z| > 2

4 O7K A 203 1 38,39,39 0.71 1 (2%) 52,60,60 1.53 5 (9%)

3 GDP A 202 2 24,30,30 1.30 4 (16%) 31,47,47 2.33 6 (19%)5 SO4 A 204 - 4,4,4 0.60 0 6,6,6 0.61 0

In the following table, the Chirals column lists the number of chiral outliers, the number of chiralcenters analysed, the number of these observed in the model and the number de�ned in theChemical Component Dictionary. Similar counts are reported in the Torsion and Rings columns.'-' means no outliers of that kind were identi�ed.

Mol Type Chain Res Link Chirals Torsions Rings4 O7K A 203 1 - 0/10/42/42 0/4/5/5


https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#rna

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#nonstandard_residues_and_ligands





Mol Type Chain Res Link Chirals Torsions Rings3 GDP A 202 2 - 0/12/32/32 0/3/3/3

All (5) bond length outliers are listed below:

Mol Chain Res Type Atoms Z Observed(Å) Ideal(Å)3 A 202 GDP O4'-C1' 2.92 1.45 1.414 A 203 O7K C-C1 -2.84 1.38 1.513 A 202 GDP O3'-C3' 2.52 1.48 1.433 A 202 GDP C2'-C3' -2.26 1.47 1.533 A 202 GDP C6-N1 2.21 1.36 1.33

All (11) bond angle outliers are listed below:

Mol Chain Res Type Atoms Z Observed(o) Ideal(o)3 A 202 GDP C5-C6-N1 -9.76 110.08 123.434 A 203 O7K C3-C4-C5 5.84 118.01 109.523 A 202 GDP C6-N1-C2 5.04 123.93 115.934 A 203 O7K C14-C15-C16 -4.86 120.83 122.484 A 203 O7K C24-N1-C5 3.95 122.10 117.893 A 202 GDP C2-N3-C4 -3.59 111.25 115.364 A 203 O7K C6-C10-C9 -2.98 118.47 121.393 A 202 GDP C4-C5-N7 -2.41 106.89 109.403 A 202 GDP C2'-C3'-C4' 2.29 107.10 102.643 A 202 GDP C6-C5-C4 2.11 122.81 120.804 A 203 O7K C6-N1-C5 -2.04 120.14 122.43

There are no chirality outliers.

There are no torsion outliers.

There are no ring outliers.

No monomer is involved in short contacts.

The following is a two-dimensional graphical depiction of Mogul quality analysis of bond lengths,bond angles, torsion angles, and ring geometry for all instances of the Ligand of Interest. Inaddition, ligands with molecular weight > 250 and outliers as shown on the validation Tables willalso be included. For torsion angles, if less then 5% of the Mogul distribution of torsion angles iswithin 10 degrees of the torsion angle in question, then that torsion angle is considered an outlier.Any bond that is central to one or more torsion angles identi�ed as an outlier by Mogul will behighlighted in the graph. For rings, the root-mean-square deviation (RMSD) between the ringin question and similar rings identi�ed by Mogul is calculated over all ring torsion angles. If theaverage RMSD is greater than 60 degrees and the minimal RMSD between the ring in question andany Mogul-identi�ed rings is also greater than 60 degrees, then that ring is considered an outlier.


The outliers are highlighted in purple. The color gray indicates Mogul did not �nd su�cientequivalents in the CSD to analyse the geometry.

Ligand O7K A 203

Bond lengths Bond angles

Torsions Rings

Ligand GDP A 202

Bond lengths Bond angles

Torsions Rings


5.7 Other polymers iO

There are no such residues in this entry.

5.8 Polymer linkage issues iO

There are no chain breaks in this entry.


https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#polymer_linkage


6 Fit of model and data iO

6.1 Protein, DNA and RNA chains iO

In the following table, the column labelled `#RSRZ> 2' contains the number (and percentage)of RSRZ outliers, followed by percent RSRZ outliers for the chain as percentile scores relative toall X-ray entries and entries of similar resolution. The OWAB column contains the minimum,median, 95th percentile and maximum values of the occupancy-weighted average B-factor perresidue. The column labelled `Q< 0.9' lists the number of (and percentage) of residues with anaverage occupancy less than 0.9.

Mol Chain Analysed <RSRZ> #RSRZ>2 OWAB(Å2) Q<0.9

1 A 170/170 (100%) 0.45 5 (2%) 51 54 3, 8, 17, 36 0

All (5) RSRZ outliers are listed below:

Mol Chain Res Type RSRZ1 A 0 GLY 3.61 A 31 GLU 2.21 A 2 THR 2.21 A 108 ASP 2.11 A 107 GLU 2.1

6.2 Non-standard residues in protein, DNA, RNA chains iO

There are no non-standard protein/DNA/RNA residues in this entry.

6.3 Carbohydrates iO

There are no monosaccharides in this entry.

6.4 Ligands iO

In the following table, the Atoms column lists the number of modelled atoms in the group and thenumber de�ned in the chemical component dictionary. The B-factors column lists the minimum,median, 95th percentile and maximum values of B factors of atoms in the group. The columnlabelled `Q< 0.9' lists the number of atoms with occupancy less than 0.9.

Mol Type Chain Res Atoms RSCC RSR B-factors(Å2) Q<0.94 O7K A 203 35/35 0.96 0.11 5,6,8,9 05 SO4 A 204 5/5 0.97 0.22 13,16,19,19 0


https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#fit_model_data

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#fit_protein_na_chains

https://www.wwpdb.org/validation/2017/XrayValidationReportHelp#fit_nonstandard_and_ligands





Mol Type Chain Res Atoms RSCC RSR B-factors(Å2) Q<0.93 GDP A 202 28/28 0.99 0.06 2,5,7,10 02 MG A 201 1/1 1.00 0.04 3,3,3,3 0

The following is a graphical depiction of the model �t to experimental electron density of allinstances of the Ligand of Interest. In addition, ligands with molecular weight > 250 and outliersas shown on the geometry validation Tables will also be included. Each �t is shown from di�erentorientation to approximate a three-dimensional view.

Electron density around O7K A 203:

2mFo-DFc (at 0.7 rmsd) in gray

mFo-DFc (at 3 rmsd) in purple (negative)

and green (positive)


Electron density around GDP A 202:

2mFo-DFc (at 0.7 rmsd) in gray

mFo-DFc (at 3 rmsd) in purple (negative)

and green (positive)

6.5 Other polymers iO

There are no such residues in this entry.


Documents

Full wwPDB X-ray Structure Validation Report i...Total C N O P 28 10 5 11 2 0 0 • Molecule 4 is pyrazinoquinolinone (three-letter code: O7K) (formula: C 25H 23ClF 2N 4O 3) (labeledas"LigandofInterest"byauthor)