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.
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) :298.000
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
56 ARG ( 58-) A
5 TYR ( 7-) A
64 PHE ( 66-) A 128 PHE ( 131-) A
12 GLU ( 14-) A 24 GLU ( 26-) A 236 GLU ( 239-) 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.
1 HIS ( 3-) A N CA 1.54 4.1 23 GLY ( 25-) A N CA 1.54 5.5 164 ILE ( 167-) A CA CB 1.63 5.1 213 ILE ( 216-) A CG1 CD1 1.67 4.0
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
| 0.993345 -0.000241 -0.001204| | -0.000241 0.994027 0.000632| | -0.001204 0.000632 0.994651|Proposed new scale matrix
| 0.023614 0.000002 0.006124| | 0.000006 0.024204 -0.000015| | 0.000017 -0.000009 0.014274|With corresponding cell
A = 42.362 B = 41.316 C = 72.396 Alpha= 89.923 Beta= 104.609 Gamma= 90.028
The CRYST1 cell dimensions
A = 42.645 B = 41.564 C = 72.750 Alpha= 90.000 Beta= 104.500 Gamma= 90.000
(Under-)estimated Z-score: 13.320
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 109.88 4.3 87 ARG ( 89-) A CB CG CD 104.86 -4.6 117 HIS ( 119-) A CG ND1 CE1 110.37 4.8 202 GLU ( 205-) A CA CB CG 104.87 -4.6
12 GLU ( 14-) A 24 GLU ( 26-) A 56 ARG ( 58-) A 236 GLU ( 239-) A
32 ASP ( 34-) A 5.10 94 HIS ( 96-) A 4.65 17 ASP ( 19-) A 4.09
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.
164 ILE ( 167-) A -2.3 81 PRO ( 83-) A -2.3 199 PRO ( 202-) A -2.1 148 GLY ( 151-) 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.
27 SER ( 29-) A PRO omega poor 55 LEU ( 57-) A omega poor 62 HIS ( 64-) A Poor phi/psi 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 240 ASP ( 243-) A Poor phi/psi 249 LYS ( 252-) A Poor phi/psi chi-1/chi-2 correlation Z-score : -0.505
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 27 SER ( 29-) A 0 48 SER ( 50-) A 0 50 ASP ( 52-) A 0 52 ALA ( 54-) A 0 60 ASN ( 62-) A 0 62 HIS ( 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 0 101 GLN ( 103-) A 0 104 GLU ( 106-) A 0And so on for a total of 119 lines.
19 PRO ( 21-) A 0.17 LOW 198 PRO ( 201-) A 0.46 HIGH 244 PRO ( 247-) A 0.06 LOW
212 PRO ( 215-) A 49.1 half-chair C-delta/C-gamma (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.
1 HIS ( 3-) A NE2 <-> 62 HIS ( 64-) A ND1 0.35 2.65 INTRA 34 HIS ( 36-) A ND1 <-> 261 HOH ( 366 ) A O 0.26 2.44 INTRA 13 HIS ( 15-) A ND1 <-> 16 LYS ( 18-) A NZ 0.21 2.79 INTRA 249 LYS ( 252-) A NZ <-> 261 HOH ( 436 ) A O 0.17 2.53 INTRA 105 HIS ( 107-) A NE2 <-> 191 TYR ( 194-) A OH 0.06 2.64 INTRA BL 115 GLU ( 117-) A OE2 <-> 117 HIS ( 119-) A NE2 0.05 2.65 INTRA BL 1 HIS ( 3-) A ND1 <-> 3 TRP ( 5-) A O 0.05 2.65 INTRA 73 ASP ( 75-) A OD1 <-> 87 ARG ( 89-) A NE 0.05 2.65 INTRA 221 LEU ( 224-) A CD1 <-> 261 HOH ( 378 ) A O 0.04 2.76 INTRA 235 GLU ( 238-) A N <-> 261 HOH ( 385 ) A O 0.02 2.68 INTRA 49 TYR ( 51-) A OH <-> 120 HIS ( 122-) A NE2 0.01 2.69 INTRA BL 83 ASP ( 85-) A C <-> 84 GLY ( 86-) A CA 0.01 2.29 INTRA B2
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.
8 HIS ( 10-) A -6.28 2 HIS ( 4-) A -5.38 98 LEU ( 100-) A -5.32 133 GLN ( 136-) A -5.10
Chain identifier: A
Note: Second generation quality Z-score plot
The second generation quality Z-score smoothed over a 10 residue window
is plotted as function of the residue number. Low areas in the plot (below
-1.3) indicate unusual packing.
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.
261 HOH ( 403 ) A O -22.42 -5.93 10.09
51 GLN ( 53-) A 175 ASN ( 178-) 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.
29 VAL ( 31-) A N 72 GLN ( 74-) A N 98 LEU ( 100-) A N 127 ASP ( 130-) A N 201 LEU ( 204-) A N 227 ASN ( 230-) A ND2 229 ASN ( 232-) A N 241 ASN ( 244-) A ND2 242 TRP ( 245-) A N 257 PHE ( 260-) A N Only metal coordination for 94 HIS ( 96-) A NE2 Only metal coordination for 117 HIS ( 119-) A ND1
The output gives the ion, the valency score for the ion itself, the valency score for the suggested alternative ion, and a series of possible comments *1 indicates that the suggested alternate atom type has been observed in the PDB file at another location in space. *2 indicates that WHAT IF thinks to have found this ion type in the crystallisation conditions as described in the REMARK 280 cards of the PDB file. *S Indicates that this ions is located at a special position (i.e. at a symmetry axis). N4 stands for NH4+.
260 NA ( 263-) A -.- -.- Too few ligands (1)
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 : -0.229 2nd generation packing quality : 0.745 Ramachandran plot appearance : -1.169 chi-1/chi-2 rotamer normality : -0.505 Backbone conformation : -1.023
Bond lengths : 1.166 Bond angles : 1.093 Omega angle restraints : 1.153 Side chain planarity : 1.822 Improper dihedral distribution : 1.526 (loose) B-factor distribution : 0.836 Inside/Outside distribution : 0.942
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 : 1.65
Structure Z-scores, positive is better than average:
1st generation packing quality : 0.0 2nd generation packing quality : -0.4 Ramachandran plot appearance : -1.2 chi-1/chi-2 rotamer normality : -0.8 Backbone conformation : -1.3
Bond lengths : 1.166 Bond angles : 1.093 Omega angle restraints : 1.153 Side chain planarity : 1.822 Improper dihedral distribution : 1.526 (loose) B-factor distribution : 0.836 Inside/Outside distribution : 0.942 ==============
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.