WHAT IF Check report

This file was created 2011-12-22 from WHAT_CHECK output by a conversion script. If you are new to WHAT_CHECK, please study the pdbreport pages. There also exists a legend to the output.

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.

Verification log for pdb1eo3.ent

Checks that need to be done early-on in validation

Warning: Ligands for which a topology was generated automatically

The topology for the ligands in the table below were determined automatically. WHAT IF uses a local copy of Daan van Aalten's Dundee PRODRG server to automatically generate topology information for ligands. For this PDB file that seems to have gone fine, but be aware that automatic topology generation is a complicated task. So, if you get messages that you fail to understand or that you believe are wrong, and one of these ligands is involved, then check the ligand topology first.

 497 TSP   (   6-)  C  -
 500 ACY   ( 601-)  A  -
 503 ACY   ( 602-)  B  -
 504 TSP   (   6-)  D  -

Non-validating, descriptive output paragraph

Note: Ramachandran plot

In this Ramachandran plot x-signs represent glycines, squares represent prolines, and plus-signs represent the other residues. If too many plus- signs fall outside the contoured areas then the molecule is poorly refined (or worse). Proline can only occur in the narrow region around phi=-60 that also falls within the other contour islands.

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

Note: Ramachandran plot

Chain identifier: B

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'.

  19 DTHY  (  11-)  D      C3'
  19 DTHY  (  11-)  D      O3'
  19 DTHY  (  11-)  D      C5'
  19 DTHY  (  11-)  D      C4'
  19 DTHY  (  11-)  D      O4'
  19 DTHY  (  11-)  D      C1'
  19 DTHY  (  11-)  D      C2'
  19 DTHY  (  11-)  D      N1
  19 DTHY  (  11-)  D      C6
  19 DTHY  (  11-)  D      C5
  19 DTHY  (  11-)  D      C4
  19 DTHY  (  11-)  D      O4
  19 DTHY  (  11-)  D      N3
  19 DTHY  (  11-)  D      C2
  19 DTHY  (  11-)  D      O2
  19 DTHY  (  11-)  D      C7
  34 GLN   (  16-)  A      CG
  34 GLN   (  16-)  A      CD
  34 GLN   (  16-)  A      OE1
  34 GLN   (  16-)  A      NE2
  35 LYS   (  17-)  A      CG
  35 LYS   (  17-)  A      CD
  35 LYS   (  17-)  A      CE
  35 LYS   (  17-)  A      NZ
  56 LYS   (  38-)  A      CG
And so on for a total of 154 lines.

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. TLS seems not mentioned in the header of the PDB file. But anyway, if WHAT IF complains about your B-factors, and 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:

Crystal temperature (K) :100.000

Note: B-factor plot

The average atomic B-factor per residue is plotted as function of the residue number.

Chain identifier: A

Note: B-factor plot

Chain identifier: B

Nomenclature related problems

Warning: Tyrosine convention problem

The tyrosine residues listed in the table below have their chi-2 not between -90.0 and 90.0

  30 TYR   (  12-)  A
  90 TYR   (  72-)  A
 141 TYR   ( 123-)  A
 154 TYR   ( 136-)  A
 251 TYR   ( 236-)  A
 320 TYR   (  61-)  B
 331 TYR   (  72-)  B
 470 TYR   ( 219-)  B

Warning: Phenylalanine convention problem

The phenylalanine residues listed in the table below have their chi-2 not between -90.0 and 90.0.

  65 PHE   (  47-)  A
 123 PHE   ( 105-)  A
 334 PHE   (  75-)  B
 450 PHE   ( 199-)  B

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.

  37 ASP   (  19-)  A
 108 ASP   (  90-)  A
 272 ASP   (  13-)  B
 278 ASP   (  19-)  B
 295 ASP   (  36-)  B

Warning: Glutamic acid convention problem

The glutamic acid residues listed in the table below have their chi-3 outside the -90.0 to 90.0 range, or their proton on OE1 instead of OE2.

  83 GLU   (  65-)  A
 173 GLU   ( 158-)  A
 235 GLU   ( 220-)  A
 304 GLU   (  45-)  B
 324 GLU   (  65-)  B
 452 GLU   ( 201-)  B

Geometric checks

Warning: Unusual bond lengths

The bond lengths listed in the table below were found to deviate more than 4 sigma from standard bond lengths (both standard values and sigmas for amino acid residues have been taken from Engh and Huber [REF], for DNA they were taken from Parkinson et al [REF]). In the table below for each unusual bond the bond length and the number of standard deviations it differs from the normal value is given.

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 DADE  (   2-)  C      C5   C6    1.37   -4.3
 147 ILE   ( 129-)  A      CA   CB    1.61    4.0

Warning: Possible cell scaling problem

Comparison of bond distances with Engh and Huber [REF] standard values for protein residues and Parkinson et al [REF] values for DNA/RNA shows a significant systematic deviation. It could be that the unit cell used in refinement was not accurate enough. The deformation matrix given below gives the deviations found: the three numbers on the diagonal represent the relative corrections needed along the A, B and C cell axis. These values are 1.000 in a normal case, but have significant deviations here (significant at the 99.99 percent confidence level)

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

 |  1.000097 -0.001390 -0.000059|
 | -0.001390  0.998115 -0.000646|
 | -0.000059 -0.000646  0.996994|
Proposed new scale matrix

 |  0.020840  0.006138  0.008838|
 |  0.000030  0.021526  0.005208|
 |  0.000001  0.000011  0.017093|
With corresponding cell

    A    =  48.005  B   =  48.453  C    =  63.638
    Alpha=  97.001  Beta= 108.922  Gamma= 106.477

The CRYST1 cell dimensions

    A    =  48.000  B   =  48.500  C    =  63.800
    Alpha=  97.000  Beta= 108.900  Gamma= 106.300

Variance: 101.154
(Under-)estimated Z-score: 7.412

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.

   4 DADE  (   5-)  C      C2'  C1'  N9  104.95   -5.8
  12 DADE  (   3-)  D      C2'  C1'  N9  120.91    4.2
  13 DGUA  (   4-)  D      O3'  C3'  C4'  93.76   -7.5
  13 DGUA  (   4-)  D      C2'  C1'  N9  123.15    5.6
  18 DTHY  (  10-)  D      C2'  C1'  N1  121.19    4.4
 208 HIS   ( 193-)  A      N    CA   C    99.38   -4.2
 318 HIS   (  59-)  B      CG   ND1  CE1 109.68    4.1
 362 THR   ( 106-)  B      N    CA   C    98.26   -4.6
 387 HIS   ( 131-)  B      CG   ND1  CE1 109.89    4.3

Error: Nomenclature error(s)

Checking for a hand-check. WHAT IF has over the course of this session already corrected the handedness of atoms in several residues. These were administrative corrections. These residues are listed here.

  37 ASP   (  19-)  A
  83 GLU   (  65-)  A
 108 ASP   (  90-)  A
 173 GLU   ( 158-)  A
 235 GLU   ( 220-)  A
 272 ASP   (  13-)  B
 278 ASP   (  19-)  B
 295 ASP   (  36-)  B
 304 GLU   (  45-)  B
 324 GLU   (  65-)  B
 452 GLU   ( 201-)  B

Warning: Chirality deviations detected

The atoms listed in the table below have an improper dihedral value that is deviating from expected values. As the improper dihedral values are all getting very close to ideal values in recent X-ray structures, and as we actually do not know how big the spread around these values should be, this check only warns for 6 sigma deviations.

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.

Please also see the previous table that lists a series of administrative chirality problems that were corrected automatically upon reading-in the PDB file.

 151 ILE   ( 133-)  A      C     -6.0    -7.86     0.03
The average deviation= 1.344

Error: Tau angle problems

The side chains of the residues listed in the table below contain a tau angle (N-Calpha-C) that was found to deviate from te expected value by more than 4.0 times the expected standard deviation. The number in the table is the number of standard deviations this RMS value deviates from the expected value.

  21 LEU   (   3-)  A    5.83
 165 LEU   ( 148-)  A    4.88
  67 ARG   (  49-)  A    4.75
 226 GLU   ( 211-)  A    4.65
 216 GLU   ( 201-)  A    4.62
 452 GLU   ( 201-)  B    4.48
 362 THR   ( 106-)  B    4.33
 331 TYR   (  72-)  B    4.24

Warning: High tau angle deviations

The RMS Z-score for the tau angles (N-Calpha-C) in the structure is too high. For well refined structures this number is expected to be near 1.0. The fact that it is higher than 1.5 worries us. However, we determined the tau normal distributions from 500 high-resolution X-ray structures, rather than from CSD data, so we cannot be 100 percent certain about these numbers.

Tau angle RMS Z-score : 1.587

Torsion-related checks

Warning: Torsion angle evaluation shows unusual residues

The residues listed in the table below contain bad or abnormal torsion angles.

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.

  91 PRO   (  73-)  A    -3.0
 332 PRO   (  73-)  B    -2.9
 353 THR   (  94-)  B    -2.6
 112 THR   (  94-)  A    -2.5
 147 ILE   ( 129-)  A    -2.5
 413 PRO   ( 162-)  B    -2.4
 308 ARG   (  49-)  B    -2.3
 114 THR   (  96-)  A    -2.3
 220 ILE   ( 205-)  A    -2.2
 301 THR   (  42-)  B    -2.1
 180 GLY   ( 165-)  A    -2.1
 443 ILE   ( 192-)  B    -2.1
 129 THR   ( 111-)  A    -2.0

Warning: Backbone evaluation reveals unusual conformations

The residues listed in the table below have abnormal backbone torsion angles.

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.

  90 TYR   (  72-)  A  PRO omega poor
 118 ASN   ( 100-)  A  Poor phi/psi
 130 SER   ( 112-)  A  Poor phi/psi
 138 ASN   ( 120-)  A  Poor phi/psi
 202 THR   ( 187-)  A  Poor phi/psi
 244 LYS   ( 229-)  A  Poor phi/psi
 259 ARG   ( 244-)  A  Poor phi/psi
 331 TYR   (  72-)  B  PRO omega poor
 368 SER   ( 112-)  B  Poor phi/psi
 376 ASN   ( 120-)  B  Poor phi/psi
 438 THR   ( 187-)  B  Poor phi/psi
 480 LYS   ( 229-)  B  Poor phi/psi
 495 ARG   ( 244-)  B  Poor phi/psi
 chi-1/chi-2 correlation Z-score : -2.279

Warning: Unusual backbone conformations

For the residues listed in the table below, the backbone formed by itself and two neighbouring residues on either side is in a conformation that is not seen very often in the database of solved protein structures. The number given in the table is the number of similar backbone conformations in the database with the same amino acid in the centre.

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 DGUA  (   4-)  C      0
   4 DADE  (   5-)  C      0
   5 DADE  (   7-)  C      0
   6 DTHY  (   8-)  C      0
   7 DCYT  (   9-)  C      0
   8 DTHY  (  10-)  C      0
   9 DTHY  (  11-)  C      0
  10 DCYT  (   1-)  D      0
  11 DADE  (   2-)  D      0
  12 DADE  (   3-)  D      0
  13 DGUA  (   4-)  D      0
  14 DADE  (   5-)  D      0
  15 DADE  (   7-)  D      0
  16 DTHY  (   8-)  D      0
  17 DCYT  (   9-)  D      0
  18 DTHY  (  10-)  D      0
  19 DTHY  (  11-)  D      0
  20 SER   (   2-)  A      0
  21 LEU   (   3-)  A      0
  35 LYS   (  17-)  A      0
  39 CYS   (  21-)  A      0
  45 GLU   (  27-)  A      0
  90 TYR   (  72-)  A      0
  92 ASP   (  74-)  A      0
  93 PHE   (  75-)  A      0
And so on for a total of 199 lines.

Warning: Omega angles too tightly restrained

The omega angles for trans-peptide bonds in a structure are expected to give a gaussian distribution with the average around +178 degrees and a standard deviation around 5.5 degrees. These expected values were obtained from very accurately determined structures. Many protein structures are too tightly restrained. This seems to be the case with the current structure too, as the observed standard deviation is below 4.0 degrees.

Standard deviation of omega values : 1.943

Warning: Backbone oxygen evaluation

The residues listed in the table below have an unusual backbone oxygen position.

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!

 180 GLY   ( 165-)  A   2.35   14

Bump checks

Error: Abnormally short interatomic distances

The pairs of atoms listed in the table below have an unusually short interactomic distance; each bump is listed in only one direction.

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.

  14 DADE  (   5-)  D      O3' <->  504 TSP   (   6-)  D      P      1.01    1.59  INTRA BL
   4 DADE  (   5-)  C      O3' <->  497 TSP   (   6-)  C      P      1.01    1.59  INTRA BL
   4 DADE  (   5-)  C      C3' <->  497 TSP   (   6-)  C      P      0.81    2.59  INTRA BL
  14 DADE  (   5-)  D      C3' <->  504 TSP   (   6-)  D      P      0.79    2.61  INTRA BL
   9 DTHY  (  11-)  C      N3  <->   11 DADE  (   2-)  D      N1     0.48    2.52  INTRA BF
  15 DADE  (   7-)  D      N1  <->  497 TSP   (   6-)  C      N3     0.36    2.64  INTRA BL
 320 TYR   (  61-)  B      CE1 <->  338 LYS   (  79-)  B      NZ     0.35    2.75  INTRA
 253 ASN   ( 238-)  A      CG  <->  257 ARG   ( 242-)  A      NH1    0.27    2.83  INTRA BF
 104 LYS   (  86-)  A      NZ  <->  145 GLN   ( 127-)  A      OE1    0.24    2.46  INTRA
  15 DADE  (   7-)  D      C2' <->   16 DTHY  (   8-)  D      O5'    0.23    2.57  INTRA
   2 DADE  (   3-)  C      N1  <->   18 DTHY  (  10-)  D      N3     0.22    2.78  INTRA BL
   8 DTHY  (  10-)  C      N3  <->   12 DADE  (   3-)  D      N1     0.22    2.78  INTRA BL
 253 ASN   ( 238-)  A      ND2 <->  257 ARG   ( 242-)  A      NH1    0.21    2.64  INTRA BF
 261 SER   (   2-)  B      N   <->  264 SER   (   5-)  B      OG     0.19    2.51  INTRA BL
  11 DADE  (   2-)  D      C1' <->  506 HOH   ( 255 )  D      O      0.18    2.62  INTRA BF
 110 LYS   (  92-)  A      NZ  <->  507 HOH   ( 694 )  A      O      0.17    2.53  INTRA
  18 DTHY  (  10-)  D      C2' <->   19 DTHY  (  11-)  D      O5'    0.17    2.63  INTRA BF
  20 SER   (   2-)  A      N   <->   23 SER   (   5-)  A      OG     0.15    2.55  INTRA BL
   5 DADE  (   7-)  C      N1  <->  504 TSP   (   6-)  D      N3     0.14    2.86  INTRA BL
  11 DADE  (   2-)  D      C2' <->   12 DADE  (   3-)  D      O5'    0.14    2.66  INTRA BF
   3 DGUA  (   4-)  C      N7  <->  436 ASN   ( 185-)  B      N      0.14    2.86  INTRA BL
  47 LYS   (  29-)  A      NZ  <->  273 GLU   (  14-)  B      OE2    0.13    2.57  INTRA BF
   1 DADE  (   2-)  C      C3' <->  474 SER   ( 223-)  B      OG     0.13    2.67  INTRA BF
  15 DADE  (   7-)  D      N3  <->  506 HOH   ( 144 )  D      O      0.13    2.57  INTRA
 133 ARG   ( 115-)  A      NH2 <->  234 TYR   ( 219-)  A      O      0.11    2.59  INTRA BL
And so on for a total of 89 lines.

Packing, accessibility and threading

Note: Inside/Outside RMS Z-score plot

The Inside/Outside distribution normality RMS Z-score over a 15 residue window is plotted as function of the residue number. High areas in the plot (above 1.5) indicate unusual inside/outside patterns.

Chain identifier: A

Note: Inside/Outside RMS Z-score plot

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.

 236 ARG   ( 221-)  A      -6.82
 195 LEU   ( 180-)  A      -6.56
 431 LEU   ( 180-)  B      -6.26
 472 ARG   ( 221-)  B      -6.14
  96 TYR   (  78-)  A      -5.55
 495 ARG   ( 244-)  B      -5.53
 415 LYS   ( 164-)  B      -5.49
 328 GLN   (  69-)  B      -5.45
 259 ARG   ( 244-)  A      -5.27
  87 GLN   (  69-)  A      -5.18
 452 GLU   ( 201-)  B      -5.12
 216 GLU   ( 201-)  A      -5.12
 179 LYS   ( 164-)  A      -5.07

Note: Quality value plot

The quality value smoothed over a 10 residue window is plotted as function of the residue number. Low areas in the plot (below -2.0) indicate unusual packing.

Chain identifier: A

Note: Quality value plot

The quality value smoothed over a 10 residue window is plotted as function of the residue number. Low areas in the plot (below -2.0) indicate unusual packing.

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.

 176 LYS   ( 161-)  A   -3.52
  86 GLN   (  68-)  A   -3.03
 327 GLN   (  68-)  B   -3.00
 400 LYS   ( 149-)  B   -2.86
 329 ASN   (  70-)  B   -2.79
 360 LYS   ( 104-)  B   -2.58
 260 LYS   ( 245-)  A   -2.55
  88 ASN   (  70-)  A   -2.54
  56 LYS   (  38-)  A   -2.54
 399 LEU   ( 148-)  B   -2.53
 116 LYS   (  98-)  A   -2.53
  35 LYS   (  17-)  A   -2.50

Warning: Abnormal packing Z-score for sequential residues

A stretch of at least four sequential residues with a 2nd generation packing Z-score below -1.75 was found. This could indicate that these residues are part of a strange loop or that the residues in this range are incomplete, but it might also be an indication of misthreading.

The table below lists the first and last residue in each stretch found, as well as the average residue Z-score of the series.

  84 PRO   (  66-)  A     -   88 ASN   (  70-)  A        -2.24
 115 ASN   (  97-)  A     -  118 ASN   ( 100-)  A        -1.50
 325 PRO   (  66-)  B     -  329 ASN   (  70-)  B        -2.27

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

Note: Second generation quality Z-score plot

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.

 507 HOH   ( 683 )  A      O    -18.37  -17.84  -34.32
 507 HOH   ( 715 )  A      O    -16.91   12.29  -23.63
 508 HOH   ( 638 )  B      O     17.13   -7.76   28.67
 508 HOH   ( 642 )  B      O     17.89  -11.78   30.75
 508 HOH   ( 692 )  B      O     21.07   19.54   -4.80
 508 HOH   ( 702 )  B      O    -24.11  -12.44   26.98

Error: Water molecules without hydrogen bonds

The water molecules listed in the table below do not form any hydrogen bonds, neither with the protein or DNA/RNA, nor with other water molecules. This is a strong indication of a refinement problem. The last number on each line is the identifier of the water molecule in the input file.

 505 HOH   (  20 )  C      O
 505 HOH   (  23 )  C      O
 505 HOH   (  30 )  C      O
 507 HOH   ( 634 )  A      O
 507 HOH   ( 635 )  A      O
 507 HOH   ( 656 )  A      O
 507 HOH   ( 705 )  A      O
 507 HOH   ( 707 )  A      O
 508 HOH   ( 698 )  B      O
ERROR. Strange cone in HB2INI
Affected atom    1 DADE (   2-) C      P
Expected ambiguity-2 in FILL1HARR 0 61 609

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.

 330 HIS   (  71-)  B

Warning: Buried unsatisfied hydrogen bond donors

The buried hydrogen bond donors listed in the table below have a hydrogen atom that is not involved in a hydrogen bond in the optimized hydrogen bond network.

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.

   5 DADE  (   7-)  C      N6
  15 DADE  (   7-)  D      N6
  38 VAL   (  20-)  A      N
  52 GLY   (  34-)  A      N
  67 ARG   (  49-)  A      NE
  86 GLN   (  68-)  A      N
  89 HIS   (  71-)  A      N
 130 SER   ( 112-)  A      OG
 139 ILE   ( 121-)  A      N
 157 THR   ( 139-)  A      N
 159 VAL   ( 141-)  A      N
 164 SER   ( 147-)  A      N
 172 ASN   ( 157-)  A      N
 178 TYR   ( 163-)  A      N
 223 SER   ( 208-)  A      N
 237 THR   ( 222-)  A      N
 247 ASN   ( 232-)  A      N
 279 VAL   (  20-)  B      N
 286 GLU   (  27-)  B      N
 292 LEU   (  33-)  B      N
 293 GLY   (  34-)  B      N
 295 ASP   (  36-)  B      N
 330 HIS   (  71-)  B      N
 343 ASN   (  84-)  B      N
 356 ASN   (  97-)  B      N
 368 SER   ( 112-)  B      OG
 377 ILE   ( 121-)  B      N
 397 VAL   ( 141-)  B      N
 414 TYR   ( 163-)  B      N
 416 GLY   ( 165-)  B      N
 426 VAL   ( 175-)  B      N
 459 SER   ( 208-)  B      N
 470 TYR   ( 219-)  B      OH
 483 ASN   ( 232-)  B      N
Only metal coordination for   63 GLU  (  45-) A      OE2

Warning: Buried unsatisfied hydrogen bond acceptors

The buried side-chain hydrogen bond acceptors listed in the table below are not involved in a hydrogen bond in the optimized hydrogen bond network.

Side-chain hydrogen bond acceptors buried inside the protein normally form hydrogen bonds within the protein. If there are any not hydrogen bonded in the optimized hydrogen bond network they will be listed here.

Waters are not listed by this option.

 208 HIS   ( 193-)  A      ND1

Warning: Unusual ion packing

We implemented the ion valence determination method of Brown and Wu [REF] similar to Nayal and Di Cera [REF]. See also Mueller, Koepke and Sheldrick [REF]. It must be stated that the validation of ions in PDB files is very difficult. Ideal ion-ligand distances often differ no more than 0.1 Angstrom, and in a 2.0 Angstrom resolution structure 0.1 Angstrom is not very much. Nayal and Di Cera showed that this method has great potential, but the method has not been validated. Part of our implementation (comparing ion types) is even fully new and despite that we see it work well in the few cases that are trivial, we must emphasize that this validation method is untested. See: swift.cmbi.ru.nl/teach/theory/ for a detailed explanation.

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+.

 499  MG   ( 502-)  A     0.56   1.12 Scores about as good as NA

Warning: Possible wrong residue type

The residues listed in the table below have a weird environment that cannot be improved by rotamer flips. This can mean one of three things, non of which WHAT CHECK really can do much about. 1) The side chain has actually another rotamer than is present in the PDB file; 2) A counter ion is present in the structure but is not given in the PDB file; 3) The residue actually is another amino acid type. The annotation 'Alt-rotamer' indicates that WHAT CHECK thinks you might want to find an alternate rotamer for this residue. The annotation 'Sym-induced' indicates that WHAT CHECK believes that symmetry contacts might have something to do with the difficulties of this residue's side chain. Determination of these two annotations is difficult, so their absence is less meaningful than their presence. The annotation Ligand-bound indicates that a ligand seems involved with this residue. In nine of ten of these cases this indicates that the ligand is causing the weird situation rather than the residue.

  37 ASP   (  19-)  A   H-bonding suggests Asn; but Alt-Rotamer; Ligand-contact
 278 ASP   (  19-)  B   H-bonding suggests Asn; but Alt-Rotamer
 479 ASP   ( 228-)  B   H-bonding suggests Asn; but Alt-Rotamer

Final summary

Note: Summary report for users of a structure

This is an overall summary of the quality of the structure as compared with current reliable structures. This summary is most useful for biologists seeking a good structure to use for modelling calculations.

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 :  -0.830
  2nd generation packing quality :  -1.910
  Ramachandran plot appearance   :  -1.645
  chi-1/chi-2 rotamer normality  :  -2.279
  Backbone conformation          :  -0.014

RMS Z-scores, should be close to 1.0:
  Bond lengths                   :   0.681
  Bond angles                    :   0.838
  Omega angle restraints         :   0.353 (tight)
  Side chain planarity           :   0.599 (tight)
  Improper dihedral distribution :   1.145
  B-factor distribution          :   0.786
  Inside/Outside distribution    :   0.986

Note: Summary report for depositors of a structure

This is an overall summary of the quality of the X-ray structure as compared with structures solved at similar resolutions. This summary can be useful for a crystallographer to see if the structure makes the best possible use of the data. Warning. This table works well for structures solved in the resolution range of the structures in the WHAT IF database, which is presently (summer 2008) mainly 1.1 - 1.3 Angstrom. The further the resolution of your file deviates from this range the more meaningless this table becomes.

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 : 2.00


Structure Z-scores, positive is better than average:

  1st generation packing quality :  -0.5
  2nd generation packing quality :  -1.3
  Ramachandran plot appearance   :  -1.0
  chi-1/chi-2 rotamer normality  :  -1.3
  Backbone conformation          :  -0.2

RMS Z-scores, should be close to 1.0:
  Bond lengths                   :   0.681
  Bond angles                    :   0.838
  Omega angle restraints         :   0.353 (tight)
  Side chain planarity           :   0.599 (tight)
  Improper dihedral distribution :   1.145
  B-factor distribution          :   0.786
  Inside/Outside distribution    :   0.986
==============

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.