Please note that you are looking at an abridged version of the output (all checks that gave normal results have been removed from this report). You can have a look at the Full report instead.
259 CO3 ( 501-) A -
Alternate atom indicators in PDB files are known to often be erroneous. It has been observed that alternate atom indicators are missing, or that there are too many of them. It is common to see that the distance between two atoms that should be covalently bound is far too big, but the distance between the alternate A of one of them and alternate B of the other is proper for a covalent bond. We have discovered many, many ways in which alternate atoms can be abused. The software tries to deal with most cases, but we know for sure that it cannot deal with all cases. If an alternate atom indicator problem is not properly solved, subsequent checks will list errors that are based on wrong coordinate combinations. So, any problem listed in this table should be solved before error messages further down in this report can be trusted.
162 ASP ( 165-) A -
In case any of these residues shows up as poor or bad in checks further down this report, please check the consistency of the alternate atoms in this residue first, correct it yourself if needed, and run the validation again.
162 ASP ( 165-) A -
In a colour picture, the residues that are part of a helix are shown in blue, strand residues in red. Preferred regions for helical residues are drawn in blue, for strand residues in red, and for all other residues in green. A full explanation of the Ramachandran plot together with a series of examples can be found at the WHAT_CHECK website.
Chain identifier: A
Coordinate problems, unexpected atoms, B-factor and occupancy checks
Warning: Missing atoms
The atoms listed in the table below are missing from the entry. If many atoms
are missing, the other checks can become less sensitive. Be aware that it
often happens that groups at the termini of DNA or RNA are really missing,
so that the absence of these atoms normally is neither an error nor the
result of poor electron density. Some of the atoms listed here might also be
listed by other checks, most noticeably by the options in the previous
section that list missing atoms in several categories. The plausible atoms
with zero occupancy are not listed here, as they already got assigned a
non-zero occupancy, and thus are no longer 'missing'.
2 HIS ( 4-) A CG 2 HIS ( 4-) A ND1 2 HIS ( 4-) A CD2 2 HIS ( 4-) A CE1 2 HIS ( 4-) A NE2 7 LYS ( 9-) A CG 7 LYS ( 9-) A CD 7 LYS ( 9-) A CE 7 LYS ( 9-) A NZ 8 HIS ( 10-) A CG 8 HIS ( 10-) A ND1 8 HIS ( 10-) A CD2 8 HIS ( 10-) A CE1 8 HIS ( 10-) A NE2 43 LYS ( 45-) A CG 43 LYS ( 45-) A CD 43 LYS ( 45-) A CE 43 LYS ( 45-) A NZ 169 LYS ( 172-) A CG 169 LYS ( 172-) A CD 169 LYS ( 172-) A CE 169 LYS ( 172-) A NZ
Obviously, the temperature at which the X-ray data was collected has some importance too:
Crystal temperature (K) :110.000
Note: B-factor plot
The average atomic B-factor per residue is plotted as function of the residue
Chain identifier: A
Nomenclature related problems
Warning: Aspartic acid convention problem
The aspartic acid residues listed in the table below have their chi-2 not
between -90.0 and 90.0, or their proton on OD1 instead of OD2.
32 ASP ( 34-) A 50 ASP ( 52-) A 83 ASP ( 85-) A 172 ASP ( 175-) A
12 GLU ( 14-) A 184 GLU ( 187-) A 218 GLU ( 221-) A
RMS Z-score for bond lengths: 0.512
RMS-deviation in bond distances: 0.010
Warning: Unusual bond angles
The bond angles listed in the table below were found to deviate more than 4
sigma from standard bond angles (both standard values and sigma for protein
residues have been taken from Engh and Huber [REF], for DNA/RNA from
Parkinson et al [REF]). In the table below for each strange angle the bond
angle and the number of standard deviations it differs from the standard
values is given. Please note that disulphide bridges are neglected. Atoms
starting with "-" belong to the previous residue in the sequence.
1 HIS ( 3-) A CG ND1 CE1 109.78 4.2 13 HIS ( 15-) A CG ND1 CE1 112.94 7.3 13 HIS ( 15-) A ND1 CE1 NE2 106.10 -4.3 15 HIS ( 17-) A CG ND1 CE1 110.38 4.8 18 PHE ( 20-) A CA CB CG 109.74 -4.1 39 ASP ( 41-) A CA CB CG 118.21 5.6 44 PRO ( 46-) A -O -C N 116.33 -4.1 56 ARG ( 58-) A CD NE CZ 129.04 4.1 81 PRO ( 83-) A -O -C N 127.90 4.2 87 ARG ( 89-) A CB CG CD 105.82 -4.1 90 GLN ( 92-) A CG CD NE2 109.77 -4.4 91 PHE ( 93-) A CA CB CG 119.39 5.6 105 HIS ( 107-) A CG ND1 CE1 109.68 4.1 120 HIS ( 122-) A CA CB CG 118.59 4.8 120 HIS ( 122-) A CB CG CD2 134.35 4.0 159 ASP ( 162-) A CA CB CG 105.98 -6.6 176 PHE ( 179-) A CA CB CG 107.76 -6.0 187 ASP ( 190-) A -O -C N 113.63 -5.9 187 ASP ( 190-) A -C N CA 133.68 6.7 187 ASP ( 190-) A CA CB CG 116.72 4.1 222 LYS ( 225-) A CA CB CG 105.26 -4.4 223 PHE ( 226-) A CA CB CG 120.34 6.5 224 ARG ( 227-) A CD NE CZ 129.66 4.4 227 ASN ( 230-) A CA CB CG 117.32 4.7
12 GLU ( 14-) A 32 ASP ( 34-) A 50 ASP ( 52-) A 83 ASP ( 85-) A 172 ASP ( 175-) A 184 GLU ( 187-) A 218 GLU ( 221-) A
These scores give an impression of how `normal' the torsion angles in protein residues are. All torsion angles except omega are used for calculating a `normality' score. Average values and standard deviations were obtained from the residues in the WHAT IF database. These are used to calculate Z-scores. A residue with a Z-score of below -2.0 is poor, and a score of less than -3.0 is worrying. For such residues more than one torsion angle is in a highly unlikely position.
199 PRO ( 202-) A -2.4 173 PHE ( 176-) A -2.2 160 VAL ( 163-) A -2.1 90 GLN ( 92-) A -2.1 81 PRO ( 83-) A -2.0
Residues with `forbidden' phi-psi combinations are listed, as well as residues with unusual omega angles (deviating by more than 3 sigma from the normal value). Please note that it is normal if about 5 percent of the residues is listed here as having unusual phi-psi combinations.
27 SER ( 29-) A PRO omega poor 55 LEU ( 57-) A omega poor 63 ALA ( 65-) A omega poor 73 ASP ( 75-) A Poor phi/psi 90 GLN ( 92-) A omega poor 109 LYS ( 111-) A Poor phi/psi 175 ASN ( 178-) A Poor phi/psi 194 SER ( 197-) A omega poor 198 PRO ( 201-) A PRO omega poor 200 LEU ( 203-) A Poor phi/psi 204 VAL ( 207-) A omega poor 212 PRO ( 215-) A omega poor 240 ASP ( 243-) A Poor phi/psi 249 LYS ( 252-) A Poor phi/psi chi-1/chi-2 correlation Z-score : -0.206
For this check, backbone conformations are compared with database structures using C-alpha superpositions with some restraints on the backbone oxygen positions.
A residue mentioned in the table can be part of a strange loop, or there might be something wrong with it or its directly surrounding residues. There are a few of these in every protein, but in any case it is worth looking at!
3 TRP ( 5-) A 0 5 TYR ( 7-) A 0 8 HIS ( 10-) A 0 17 ASP ( 19-) A 0 18 PHE ( 20-) A 0 22 LYS ( 24-) A 0 25 ARG ( 27-) A 0 26 GLN ( 28-) A 0 27 SER ( 29-) A 0 36 ALA ( 38-) A 0 48 SER ( 50-) A 0 50 ASP ( 52-) A 0 52 ALA ( 54-) A 0 60 ASN ( 62-) A 0 62 CYS ( 64-) A 0 70 ASP ( 72-) A 0 71 SER ( 73-) A 0 73 ASP ( 75-) A 0 74 LYS ( 76-) A 0 75 ALA ( 77-) A 0 78 LYS ( 80-) A 0 81 PRO ( 83-) A 0 83 ASP ( 85-) A 0 90 GLN ( 92-) A 0 97 SER ( 99-) A 0And so on for a total of 118 lines.
19 PRO ( 21-) A 0.17 LOW 135 PRO ( 138-) A 0.17 LOW 244 PRO ( 247-) A 0.17 LOW
The contact distances of all atom pairs have been checked. Two atoms are said to `bump' if they are closer than the sum of their Van der Waals radii minus 0.40 Angstrom. For hydrogen bonded pairs a tolerance of 0.55 Angstrom is used. The first number in the table tells you how much shorter that specific contact is than the acceptable limit. The second distance is the distance between the centres of the two atoms. Although we believe that two water atoms at 2.4 A distance are too close, we only report water pairs that are closer than this rather short distance.
The last text-item on each line represents the status of the atom pair. If the final column contains the text 'HB', the bump criterion was relaxed because there could be a hydrogen bond. Similarly relaxed criteria are used for 1-3 and 1-4 interactions (listed as 'B2' and 'B3', respectively). BL indicates that the B-factors of the clashing atoms have a low B-factor thereby making this clash even more worrisome. INTRA and INTER indicate whether the clashes are between atoms in the same asymmetric unit, or atoms in symmetry related asymmetric units, respectively.
25 ARG ( 27-) A A NH1 <-> 260 HOH (1175 ) A O 0.22 2.48 INTRA 1 HIS ( 3-) A NE2 <-> 62 CYS ( 64-) A A SG 0.21 3.09 INTRA BF 25 ARG ( 27-) A A NH2 <-> 260 HOH (1175 ) A O 0.21 2.49 INTRA 249 LYS ( 252-) A NZ <-> 260 HOH (1109 ) A O 0.21 2.49 INTRA 124 LYS ( 127-) A NZ <-> 260 HOH (1149 ) A O 0.17 2.53 INTRA BF 105 HIS ( 107-) A NE2 <-> 191 TYR ( 194-) A OH 0.14 2.56 INTRA BL 3 TRP ( 5-) A CH2 <-> 62 CYS ( 64-) A A SG 0.12 3.28 INTRA 159 ASP ( 162-) A OD2 <-> 260 HOH (1173 ) A O 0.11 2.29 INTRA 1 HIS ( 3-) A CE1 <-> 62 CYS ( 64-) A A SG 0.09 3.31 INTRA BF 165 LYS ( 168-) A NZ <-> 235 GLU ( 238-) A OE2 0.05 2.65 INTRA BF 115 GLU ( 117-) A OE2 <-> 117 HIS ( 119-) A NE2 0.02 2.68 INTRA BL 175 ASN ( 178-) A N <-> 260 HOH (1022 ) A O 0.02 2.68 INTRA 43 LYS ( 45-) A O <-> 260 HOH (1036 ) A O 0.02 2.38 INTRA 1 HIS ( 3-) A CD2 <-> 3 TRP ( 5-) A O 0.01 2.79 INTRA 25 ARG ( 27-) A A CZ <-> 260 HOH (1175 ) A O 0.01 2.79 INTRA 22 LYS ( 24-) A CE <-> 260 HOH (1169 ) A O 0.01 2.79 INTRA
Chain identifier: A
Warning: Abnormal packing environment for some residues
The residues listed in the table below have an unusual packing environment.
The packing environment of the residues is compared with the average packing environment for all residues of the same type in good PDB files. A low packing score can indicate one of several things: Poor packing, misthreading of the sequence through the density, crystal contacts, contacts with a co-factor, or the residue is part of the active site. It is not uncommon to see a few of these, but in any case this requires further inspection of the residue.
98 LEU ( 100-) A -5.30
Chain identifier: A
Warning: Low packing Z-score for some residues
The residues listed in the table below have an unusual packing
environment according to the 2nd generation packing check. The score
listed in the table is a packing normality Z-score: positive means
better than average, negative means worse than average. Only residues
scoring less than -2.50 are listed here. These are the unusual
residues in the structure, so it will be interesting to take a
special look at them.
43 LYS ( 45-) A -2.94 8 HIS ( 10-) A -2.89 2 HIS ( 4-) A -2.69
Chain identifier: A
Water, ion, and hydrogenbond related checks
Warning: Water molecules need moving
The water molecules listed in the table below were found to be significantly
closer to a symmetry related non-water molecule than to the ones given in the
coordinate file. For optimal viewing convenience revised coordinates for
these water molecules should be given.
The number in brackets is the identifier of the water molecule in the input file. Suggested coordinates are also given in the table. Please note that alternative conformations for protein residues are not taken into account for this calculation. If you are using WHAT IF / WHAT-CHECK interactively, then the moved waters can be found in PDB format in the file: MOVEDH2O.pdb.
260 HOH (1151 ) A O 12.05 8.04 28.99 260 HOH (1154 ) A O 28.55 -1.75 -10.61
260 HOH (1125 ) A O 260 HOH (1172 ) A O Metal-coordinating Histidine residue 92 fixed to 1 Metal-coordinating Histidine residue 94 fixed to 1 Metal-coordinating Histidine residue 117 fixed to 1
1 HIS ( 3-) A 133 GLN ( 136-) A
Hydrogen bond donors that are buried inside the protein normally use all of their hydrogens to form hydrogen bonds within the protein. If there are any non hydrogen bonded buried hydrogen bond donors in the structure they will be listed here. In very good structures the number of listed atoms will tend to zero.
Waters are not listed by this option.
3 TRP ( 5-) A N 29 VAL ( 31-) A N 98 LEU ( 100-) A N 162 ASP ( 165-) A N 197 THR ( 200-) A N 201 LEU ( 204-) A N 227 ASN ( 230-) A ND2 242 TRP ( 245-) A N Only metal coordination for 94 HIS ( 96-) A NE2 Only metal coordination for 117 HIS ( 119-) A ND1
The score listed is the valency score. This number should be close to (preferably a bit above) 1.0 for the suggested ion to be a likely alternative for the water molecule. Ions listed in brackets are good alternate choices. *1 indicates that the suggested ion-type has been observed elsewhere in the PDB file too. *2 indicates that the suggested ion-type has been observed in the REMARK 280 cards of the PDB file. Ion-B and ION-B indicate that the B-factor of this water is high, or very high, respectively. H2O-B indicates that the B-factors of atoms that surround this water/ion are suspicious. See: swift.cmbi.ru.nl/teach/theory/ for a detailed explanation.
260 HOH (1077 ) A O 0.96 K 4 ION-B H2O-B
159 ASP ( 162-) A H-bonding suggests Asn; but Alt-Rotamer
The second part of the table mostly gives an impression of how well the model conforms to common refinement restraint values. The first part of the table shows a number of global quality indicators.
Structure Z-scores, positive is better than average:
1st generation packing quality : 0.326 2nd generation packing quality : 0.744 Ramachandran plot appearance : -1.090 chi-1/chi-2 rotamer normality : -0.206 Backbone conformation : -1.043
Bond lengths : 0.512 (tight) Bond angles : 1.221 Omega angle restraints : 1.128 Side chain planarity : 0.621 (tight) Improper dihedral distribution : 0.738 B-factor distribution : 1.060 Inside/Outside distribution : 0.962
The second part of the table mostly gives an impression of how well the model conforms to common refinement restraint values. The first part of the table shows a number of global quality indicators, which have been calibrated against structures of similar resolution.
Resolution found in PDB file : 1.40
Structure Z-scores, positive is better than average:
1st generation packing quality : 0.5 2nd generation packing quality : -0.2 Ramachandran plot appearance : -1.5 chi-1/chi-2 rotamer normality : -0.8 Backbone conformation : -1.4
Bond lengths : 0.512 (tight) Bond angles : 1.221 Omega angle restraints : 1.128 Side chain planarity : 0.621 (tight) Improper dihedral distribution : 0.738 B-factor distribution : 1.060 Inside/Outside distribution : 0.962 ==============
WHAT IF G.Vriend, WHAT IF: a molecular modelling and drug design program, J. Mol. Graph. 8, 52--56 (1990). WHAT_CHECK (verification routines from WHAT IF) R.W.W.Hooft, G.Vriend, C.Sander and E.E.Abola, Errors in protein structures Nature 381, 272 (1996). (see also http://swift.cmbi.ru.nl/gv/whatcheck for a course and extra inform Bond lengths and angles, protein residues R.Engh and R.Huber, Accurate bond and angle parameters for X-ray protein structure refinement, Acta Crystallogr. A47, 392--400 (1991). Bond lengths and angles, DNA/RNA G.Parkinson, J.Voitechovsky, L.Clowney, A.T.Bruenger and H.Berman, New parameters for the refinement of nucleic acid-containing structures Acta Crystallogr. D52, 57--64 (1996). DSSP W.Kabsch and C.Sander, Dictionary of protein secondary structure: pattern recognition of hydrogen bond and geometrical features Biopolymers 22, 2577--2637 (1983). Hydrogen bond networks R.W.W.Hooft, C.Sander and G.Vriend, Positioning hydrogen atoms by optimizing hydrogen bond networks in protein structures PROTEINS, 26, 363--376 (1996). Matthews' Coefficient B.W.Matthews Solvent content of Protein Crystals J. Mol. Biol. 33, 491--497 (1968). Protein side chain planarity R.W.W. Hooft, C. Sander and G. Vriend, Verification of protein structures: side-chain planarity J. Appl. Cryst. 29, 714--716 (1996). Puckering parameters D.Cremer and J.A.Pople, A general definition of ring puckering coordinates J. Am. Chem. Soc. 97, 1354--1358 (1975). Quality Control G.Vriend and C.Sander, Quality control of protein models: directional atomic contact analysis, J. Appl. Cryst. 26, 47--60 (1993). Ramachandran plot G.N.Ramachandran, C.Ramakrishnan and V.Sasisekharan, Stereochemistry of Polypeptide Chain Conformations J. Mol. Biol. 7, 95--99 (1963). Symmetry Checks R.W.W.Hooft, C.Sander and G.Vriend, Reconstruction of symmetry related molecules from protein data bank (PDB) files J. Appl. Cryst. 27, 1006--1009 (1994). Ion Checks I.D.Brown and K.K.Wu, Empirical Parameters for Calculating Cation-Oxygen Bond Valences Acta Cryst. B32, 1957--1959 (1975). M.Nayal and E.Di Cera, Valence Screening of Water in Protein Crystals Reveals Potential Na+ Binding Sites J.Mol.Biol. 256 228--234 (1996). P.Mueller, S.Koepke and G.M.Sheldrick, Is the bond-valence method able to identify metal atoms in protein structures? Acta Cryst. D 59 32--37 (2003). Checking checks K.Wilson, C.Sander, R.W.W.Hooft, G.Vriend, et al. Who checks the checkers J.Mol.Biol. (1998) 276,417-436.