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: B
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) :287.000
Warning: More than 5 percent of buried atoms has low B-factor
For normal protein structures, no more than about 1 percent of the B factors
of buried atoms is below 5.0. The fact that this value is much higher in the
current structure could be a signal that the B-factors were restraints or
constraints to too-low values, misuse of B-factor field in the PDB file, or
a TLS/scaling problem. If the average B factor is low too, it is probably a
low temperature structure determination.
Percentage of buried atoms with B less than 5 : 35.45
Note: B-factor plot
The average atomic B-factor per residue is plotted as function of the residue
Chain identifier: B
Nomenclature related problems
Warning: Phenylalanine convention problem
The phenylalanine residues listed in the table below have their chi-2 not
between -90.0 and 90.0.
40 PHE ( 36-) B 50 PHE ( 46-) B 87 PHE ( 83-) B 133 PHE ( 129-) B
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.
4 HIS ( 0-) B CB CG 1.43 -4.4 5 HIS ( 1-) B CB CG 1.37 -8.7 86 LYS ( 82-) B CA CB 1.44 -4.7 86 LYS ( 82-) B CB CG 0.93 -19.7 98 GLY ( 94-) B CA C 1.45 -4.3 137 LYS ( 133-) B CA CB 1.62 4.7 138 GLU ( 134-) B CB CG 1.38 -4.6 146 MET ( 142-) B CB CG 1.37 -4.8 146 MET ( 142-) B CG SD 2.06 7.7 147 GLU ( 143-) B CB CG 1.40 -4.1 158 LYS ( 154-) B CD CE 1.85 11.0 158 LYS ( 154-) B CE NZ 1.29 -6.7 167 GLN ( 163-) B CA C 1.44 -4.2
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
| 1.005157 -0.002205 -0.000385| | -0.002205 1.002845 0.001165| | -0.000385 0.001165 1.002687|Proposed new scale matrix
| 0.023563 0.000052 0.000009| | 0.000042 0.019029 -0.000022| | 0.000004 -0.000013 0.011176|With corresponding cell
A = 42.440 B = 52.552 C = 89.478 Alpha= 89.867 Beta= 90.044 Gamma= 90.252
The CRYST1 cell dimensions
A = 42.222 B = 52.404 C = 89.236 Alpha= 90.000 Beta= 90.000 Gamma= 90.000
(Under-)estimated Z-score: 6.687
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.
5 HIS ( 1-) B CA CB CG 104.49 -9.3 12 PHE ( 8-) B CA CB CG 119.18 5.4 96 HIS ( 92-) B CG ND1 CE1 109.63 4.0 130 HIS ( 126-) B CB CG ND1 131.50 6.6 130 HIS ( 126-) B CB CG CD2 122.85 -4.8 135 LYS ( 131-) B CD CE NZ 95.02 -5.3 138 GLU ( 134-) B CG CD OE2 128.23 4.3 138 GLU ( 134-) B CG CD OE1 107.58 -4.7 146 MET ( 142-) B C CA CB 120.53 5.5 146 MET ( 142-) B CA CB CG 104.80 -4.7 158 LYS ( 154-) B CD CE NZ 66.65 -14.1
Improper dihedrals are a measure of the chirality/planarity of the structure at a specific atom. Values around -35 or +35 are expected for chiral atoms, and values around 0 for planar atoms. Planar side chains are left out of the calculations, these are better handled by the planarity checks.
Three numbers are given for each atom in the table. The first is the Z-score for the improper dihedral. The second number is the measured improper dihedral. The third number is the expected value for this atom type. A final column contains an extra warning if the chirality for an atom is opposite to the expected value.
137 LYS ( 133-) B C -6.5 -9.74 0.11 The average deviation= 1.574
84 GLY ( 80-) B 4.35
5 HIS ( 1-) B 12.41 31 ASP ( 27-) B 5.38 50 PHE ( 46-) B 4.38 3 HIS ( -1-) B 4.23
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.
74 HIS ( 70-) B -2.3 65 MET ( 61-) B -2.2 33 VAL ( 29-) B -2.1 113 GLY ( 109-) B -2.1 64 PHE ( 60-) B -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.
11 PHE ( 7-) B omega poor 51 GLY ( 47-) B omega poor 56 CYS ( 52-) B omega poor 74 HIS ( 70-) B Poor phi/psi 88 GLU ( 84-) B omega poor 110 ASN ( 106-) B Poor phi/psi chi-1/chi-2 correlation Z-score : 0.297
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!
5 HIS ( 1-) B 0 16 VAL ( 12-) B 0 20 PRO ( 16-) B 0 29 PHE ( 25-) B 0 30 ALA ( 26-) B 0 32 LYS ( 28-) B 0 33 VAL ( 29-) B 0 47 GLU ( 43-) B 0 48 LYS ( 44-) B 0 50 PHE ( 46-) B 0 52 TYR ( 48-) B 0 53 LYS ( 49-) B 0 57 PHE ( 53-) B 0 58 HIS ( 54-) B 0 62 PRO ( 58-) B 0 64 PHE ( 60-) B 0 65 MET ( 61-) B 0 72 THR ( 68-) B 0 73 ARG ( 69-) B 0 74 HIS ( 70-) B 0 75 ASN ( 71-) B 0 77 THR ( 73-) B 0 83 TYR ( 79-) B 0 85 GLU ( 81-) B 0 86 LYS ( 82-) B 0And so on for a total of 80 lines.
For each of the residues in the structure, a search was performed to find 5-residue stretches in the WHAT IF database with superposable C-alpha coordinates, and some restraining on the neighbouring backbone oxygens.
In the following table the RMS distance between the backbone oxygen positions of these matching structures in the database and the position of the backbone oxygen atom in the current residue is given. If this number is larger than 1.5 a significant number of structures in the database show an alternative position for the backbone oxygen. If the number is larger than 2.0 most matching backbone fragments in the database have the peptide plane flipped. A manual check needs to be performed to assess whether the experimental data can support that alternative as well. The number in the last column is the number of database hits (maximum 80) used in the calculation. It is "normal" that some glycine residues show up in this list, but they are still worth checking!
84 GLY ( 80-) B 2.13 11
8 PRO ( 4-) B 0.04 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.
169 HOH (2336 ) B O <-> 169 HOH (2337 ) B O 1.79 0.41 INTRA 169 HOH (2303 ) B O <-> 169 HOH (2304 ) B O 1.78 0.42 INTRA 169 HOH (2222 ) B O <-> 169 HOH (2223 ) B O 1.78 0.42 INTRA 169 HOH (2154 ) B O <-> 169 HOH (2155 ) B O 1.73 0.47 INTRA 169 HOH (2272 ) B O <-> 169 HOH (2338 ) B O 1.72 0.48 INTRA 169 HOH (2424 ) B O <-> 169 HOH (2426 ) B O 1.68 0.52 INTRA 169 HOH (2235 ) B O <-> 169 HOH (2237 ) B O 1.68 0.52 INTRA 169 HOH (2316 ) B O <-> 169 HOH (2341 ) B O 1.66 0.54 INTRA 169 HOH (2072 ) B O <-> 169 HOH (2073 ) B O 1.64 0.56 INTRA 169 HOH (2381 ) B O <-> 169 HOH (2388 ) B O 1.61 0.59 INTRA 169 HOH (2171 ) B O <-> 169 HOH (2172 ) B O 1.60 0.60 INTRA 169 HOH (2290 ) B O <-> 169 HOH (2291 ) B O 1.55 0.65 INTRA 169 HOH (2160 ) B O <-> 169 HOH (2162 ) B O 1.52 0.68 INTRA 169 HOH (2177 ) B O <-> 169 HOH (2186 ) B O 1.49 0.71 INTRA 169 HOH (2359 ) B O <-> 169 HOH (2360 ) B O 1.49 0.71 INTRA 169 HOH (2069 ) B O <-> 169 HOH (2202 ) B O 1.49 0.71 INTRA 169 HOH (2278 ) B O <-> 169 HOH (2279 ) B O 1.42 0.78 INTRA 169 HOH (2153 ) B O <-> 169 HOH (2156 ) B O 1.41 0.79 INTRA 169 HOH (2298 ) B O <-> 169 HOH (2364 ) B O 1.39 0.81 INTRA 169 HOH (2415 ) B O <-> 169 HOH (2417 ) B O 1.37 0.83 INTRA 169 HOH (2140 ) B O <-> 169 HOH (2141 ) B O 1.35 0.85 INTRA 169 HOH (2318 ) B O <-> 169 HOH (2319 ) B O 1.34 0.86 INTRA 169 HOH (2055 ) B O <-> 169 HOH (2443 ) B O 1.30 0.90 INTRA 169 HOH (2306 ) B O <-> 169 HOH (2308 ) B O 1.29 0.91 INTRA 169 HOH (2402 ) B O <-> 169 HOH (2404 ) B O 1.27 0.93 INTRAAnd so on for a total of 121 lines.
Chain identifier: B
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.
152 ARG ( 148-) B -7.97 148 ARG ( 144-) B -5.58 2 HIS ( -2-) B -5.16 5 HIS ( 1-) B -5.14 6 VAL ( 2-) B -5.13
The table below lists the first and last residue in each stretch found, as well as the average residue score of the series.
2 HIS ( -2-) B 6 - VAL 2- ( B) -4.74
Chain identifier: B
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.
91 ASN ( 87-) B -3.36 124 LYS ( 120-) B -2.54
The table below lists the first and last residue in each stretch found, as well as the average residue Z-score of the series.
3 HIS ( -1-) B - 6 VAL ( 2-) B -1.67 90 GLU ( 86-) B - 93 ILE ( 89-) B -2.02 123 THR ( 119-) B - 126 LEU ( 122-) B -1.86
Chain identifier: B
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.
169 HOH (2048 ) B O 16.72 -1.36 -30.86 169 HOH (2114 ) B O -6.83 -2.78 -32.80 169 HOH (2158 ) B O 20.66 -7.29 -0.60 169 HOH (2205 ) B O 28.04 5.93 -15.44 169 HOH (2220 ) B O 21.81 -11.46 -21.22
169 HOH (2048 ) B O 169 HOH (2112 ) B O 169 HOH (2121 ) B O ERROR. No convergence in HB2STD Old,New value: 707.769 707.780
3 HIS ( -1-) B 4 HIS ( 0-) B 5 HIS ( 1-) B 110 ASN ( 106-) B 130 HIS ( 126-) B
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.
91 ASN ( 87-) B N 121 ALA ( 117-) B N 125 TRP ( 121-) B N 158 LYS ( 154-) B N
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.
169 HOH (2080 ) B O 1.12 K 5 ION-B 169 HOH (2117 ) B O 1.07 K 4 169 HOH (2118 ) B O 0.86 K 5 Ion-B 169 HOH (2164 ) B O 0.99 K 6 169 HOH (2289 ) B O 0.97 K 4 169 HOH (2375 ) B O 1.08 CA 7 (or NA) Ion-B 169 HOH (2384 ) B O 1.14 CA 8 (or NA) Ion-B 169 HOH (2435 ) B O 1.05 MG 7 ION-B
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 : -1.106 2nd generation packing quality : -1.973 Ramachandran plot appearance : 0.298 chi-1/chi-2 rotamer normality : 0.297 Backbone conformation : -0.468
Bond lengths : 1.162 Bond angles : 1.166 Omega angle restraints : 1.105 Side chain planarity : 2.478 (loose) Improper dihedral distribution : 1.409 Inside/Outside distribution : 0.949
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 : 1.20
Structure Z-scores, positive is better than average:
1st generation packing quality : -0.6 2nd generation packing quality : -1.6 Ramachandran plot appearance : -0.2 chi-1/chi-2 rotamer normality : -0.2 Backbone conformation : -0.7
Bond lengths : 1.162 Bond angles : 1.166 Omega angle restraints : 1.105 Side chain planarity : 2.478 (loose) Improper dihedral distribution : 1.409 Inside/Outside distribution : 0.949 ==============
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.