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.
148 4G5 (1165-) A - 149 12P (1164-) 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: What type of B-factor?
WHAT IF does not yet know well how to cope with B-factors in case TLS has
been used. It simply assumes that the B-factor listed on the ATOM and HETATM
cards are the total B-factors. When TLS refinement is used that assumption
sometimes is not correct. The header of the PDB file states that TLS groups
were used. So, if WHAT IF complains about your B-factors, while you think
that they are OK, then check for TLS related B-factor problems first.
Obviously, the temperature at which the X-ray data was collected has some importance too:
Number of TLS groups mentione in PDB file header: 0
Crystal temperature (K) :277.000
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: Arginine nomenclature problem
The arginine residues listed in the table below have their N-H-1 and N-H-2
125 ARG ( 142-) A
17 TYR ( 23-) A 75 TYR ( 92-) A
86 PHE ( 103-) A 93 PHE ( 110-) A 117 PHE ( 134-) A
85 ASP ( 102-) A 95 ASP ( 112-) A
34 GLU ( 51-) A 59 GLU ( 76-) A 67 GLU ( 84-) A 70 GLU ( 87-) A 84 GLU ( 101-) A
Atom names starting with "-" belong to the previous residue in the chain. If the second atom name is "-SG*", the disulphide bridge has a deviating length.
29 GLU ( 35-) A CD OE2 1.33 4.1
There are a number of different possible causes for the discrepancy. First the cell used in refinement can be different from the best cell calculated. Second, the value of the wavelength used for a synchrotron data set can be miscalibrated. Finally, the discrepancy can be caused by a dataset that has not been corrected for significant anisotropic thermal motion.
Please note that the proposed scale matrix has NOT been restrained to obey the space group symmetry. This is done on purpose. The distortions can give you an indication of the accuracy of the determination.
If you intend to use the result of this check to change the cell dimension of your crystal, please read the extensive literature on this topic first. This check depends on the wavelength, the cell dimensions, and on the standard bond lengths and bond angles used by your refinement software.
Unit Cell deformation matrix
| 0.992277 -0.001818 0.000099| | -0.001818 0.993326 0.001335| | 0.000099 0.001335 0.993315|Proposed new scale matrix
| 0.014756 0.008529 -0.000013| | 0.000031 0.017004 -0.000023| | -0.000001 -0.000017 0.012687|With corresponding cell
A = 67.839 B = 68.000 C = 78.822 Alpha= 89.872 Beta= 89.989 Gamma= 120.131
The CRYST1 cell dimensions
A = 68.367 B = 68.367 C = 79.351 Alpha= 90.000 Beta= 90.000 Gamma= 120.000
(Under-)estimated Z-score: 11.714
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.
47 HIS ( 64-) A CG ND1 CE1 109.87 4.3 140 HIS ( 157-) A CG ND1 CE1 110.02 4.4
34 GLU ( 51-) A 59 GLU ( 76-) A 67 GLU ( 84-) A 70 GLU ( 87-) A 84 GLU ( 101-) A 85 ASP ( 102-) A 95 ASP ( 112-) A 125 ARG ( 142-) A
115 LYS ( 132-) A 4.67
118 GLU ( 135-) A 5.54
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.
42 HIS ( 59-) A -2.1
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.
12 SER ( 18-) A omega poor 13 SER ( 19-) A omega poor 24 ASN ( 30-) A Poor phi/psi 32 SER ( 38-) A omega poor 40 CYS ( 57-) A omega poor 53 PRO ( 70-) A Poor phi/psi 92 GLN ( 109-) A omega poor 102 ARG ( 119-) A Poor phi/psi 109 SER ( 126-) A omega poor 130 SER ( 147-) A omega poor 140 HIS ( 157-) A omega poor chi-1/chi-2 correlation Z-score : -1.210
It is not necessarily an error if a few residues have rotamer values below 0.3, but careful inspection of all residues with these low values could be worth it.
121 SER ( 138-) A 0.38
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 PRO ( 9-) A 0 5 TRP ( 11-) A 0 6 GLU ( 12-) A 0 13 SER ( 19-) A 0 16 VAL ( 22-) A 0 21 HIS ( 27-) A 0 24 ASN ( 30-) A 0 31 PRO ( 37-) A 0 32 SER ( 38-) A 0 33 GLY ( 39-) A 0 34 GLU ( 51-) A 0 35 PRO ( 52-) A 0 36 ALA ( 53-) A 0 47 HIS ( 64-) A 0 52 ARG ( 69-) A 0 53 PRO ( 70-) A 0 54 SER ( 71-) A 0 57 ARG ( 74-) A 0 58 GLN ( 75-) A 0 62 THR ( 79-) A 0 81 SER ( 98-) A 0 84 GLU ( 101-) A 0 102 ARG ( 119-) A 0 105 LEU ( 122-) A 0 107 ALA ( 124-) A 0And so on for a total of 62 lines.
2 PRO ( 8-) A 41.6 envelop C-delta (36 degrees) 53 PRO ( 70-) A -52.3 half-chair C-beta/C-alpha (-54 degrees)
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.
15 ARG ( 21-) A NH2 <-> 150 HOH (2008 ) A O 0.49 2.21 INTRA BF 15 ARG ( 21-) A NE <-> 150 HOH (2009 ) A O 0.39 2.31 INTRA BF 96 CYS ( 113-) A SG <-> 148 4G5 (1165-) A N4 0.24 3.06 INTRA BL 92 GLN ( 109-) A NE2 <-> 150 HOH (2079 ) A O 0.18 2.52 INTRA BL 84 GLU ( 101-) A OE2 <-> 150 HOH (2074 ) A O 0.18 2.22 INTRA BF 150 HOH (2008 ) A O <-> 150 HOH (2010 ) A O 0.15 2.05 INTRA 42 HIS ( 59-) A ND1 <-> 140 HIS ( 157-) A ND1 0.14 2.86 INTRA BL 15 ARG ( 21-) A NH2 <-> 28 TRP ( 34-) A CD2 0.12 2.98 INTRA 115 LYS ( 132-) A CE <-> 150 HOH (2095 ) A O 0.12 2.68 INTRA BF 59 GLU ( 76-) A OE2 <-> 150 HOH (2045 ) A O 0.12 2.28 INTRA BF 33 GLY ( 39-) A N <-> 150 HOH (2028 ) A O 0.11 2.59 INTRA BF 65 LYS ( 82-) A NZ <-> 150 HOH (2056 ) A O 0.10 2.60 INTRA 24 ASN ( 30-) A ND2 <-> 150 HOH (2018 ) A O 0.09 2.61 INTRA BL 33 GLY ( 39-) A CA <-> 150 HOH (2026 ) A O 0.08 2.72 INTRA BF 60 LYS ( 77-) A NZ <-> 150 HOH (2050 ) A O 0.06 2.64 INTRA BF 84 GLU ( 101-) A OE1 <-> 150 HOH (2073 ) A O 0.05 2.35 INTRA BF 114 GLN ( 131-) A NE2 <-> 150 HOH (2090 ) A O 0.05 2.65 INTRA BF 130 SER ( 147-) A N <-> 142 ILE ( 159-) A O 0.03 2.67 INTRA BL 148 4G5 (1165-) A C13 <-> 150 HOH (2091 ) A O 0.03 2.77 INTRA 119 ASP ( 136-) A OD1 <-> 150 HOH (2095 ) A O 0.01 2.39 INTRA 127 GLY ( 144-) A N <-> 144 ARG ( 161-) A O 0.01 2.69 INTRA BL
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.
51 ARG ( 68-) A -7.32 57 ARG ( 74-) A -5.73 52 ARG ( 69-) A -5.56 110 ARG ( 127-) A -5.31 92 GLN ( 109-) A -5.29 11 ARG ( 17-) A -5.23
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.
99 ALA ( 116-) A -2.51
Chain identifier: A
Water, ion, and hydrogenbond related checks
Error: HIS, ASN, GLN side chain flips
Listed here are Histidine, Asparagine or Glutamine residues for
which the orientation determined from hydrogen bonding analysis are
different from the assignment given in the input. Either they could
form energetically more favourable hydrogen bonds if the terminal
group was rotated by 180 degrees, or there is no assignment in the
input file (atom type 'A') but an assignment could be made. Be aware,
though, that if the topology could not be determined for one or more
ligands, then this option will make errors.
24 ASN ( 30-) A 92 GLN ( 109-) 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.
15 ARG ( 21-) A N 20 ASN ( 26-) A ND2 26 SER ( 32-) A N 46 LYS ( 63-) A NZ 125 ARG ( 142-) A NH2
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.
150 HOH (2020 ) A O 0.85 K 4 150 HOH (2072 ) A O 0.97 NA 4 150 HOH (2088 ) A O 1.05 K 5
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 : -1.027 2nd generation packing quality : 0.154 Ramachandran plot appearance : 0.338 chi-1/chi-2 rotamer normality : -1.210 Backbone conformation : -0.255
Bond lengths : 1.050 Bond angles : 0.972 Omega angle restraints : 1.265 Side chain planarity : 1.502 Improper dihedral distribution : 1.184 B-factor distribution : 0.617 Inside/Outside distribution : 0.990
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 : 1.90
Structure Z-scores, positive is better than average:
1st generation packing quality : -0.6 2nd generation packing quality : -0.2 Ramachandran plot appearance : 0.8 chi-1/chi-2 rotamer normality : -0.6 Backbone conformation : -0.6
Bond lengths : 1.050 Bond angles : 0.972 Omega angle restraints : 1.265 Side chain planarity : 1.502 Improper dihedral distribution : 1.184 B-factor distribution : 0.617 Inside/Outside distribution : 0.990 ==============
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.