WHAT IF Check report

This file was created 2004-09-28 from WHAT_CHECK output by a conversion script. If you are new to WHAT_CHECK, please read the introduction 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 103L

Nomenclature related problems

Warning: Chirality deviations detected

The atoms listed in the table below have an improper dihedral value that is deviating from expected values.

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.

  20 ASP  (  20 )      CA        -5.2     22.1     34.0
  90 ALA  (  93 )      CA        -4.1     26.8     34.4
 121 LYS  ( 124 )      CA        -4.5     24.9     34.3
 123 TRP  ( 126 )      CA        -4.9     22.4     34.3

Geometric checks

Warning: C-terminal oxygen atoms missing

The C-atoms listed in the table below belong to a C-terminal residue in a protein chain, but the C-terminal oxygen ("O2" or "OXT") that it should be bound to was not found.

  34 THR  (  34 )      C
 159 LYS  ( 162 )      C

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.

  20 ASP  (  20 )        CA   CB   CG  117.628   5.0
  25 TYR  (  25 )        C    CA   CB  118.406   4.4
  31 HIS  (  31 )        CA   CB   CG  108.660  -5.1
  50 ASN  (  53 )        CA   CB   CG  107.674  -4.9
 111 PHE  ( 114 )        CA   CB   CG  117.935   4.1

Warning: Unusual PRO puckering amplitudes

The proline residues listed in the table below have a puckering amplitude that is outside of normal ranges. Puckering parameters were calculated by the method of Cremer and Pople [REF]. Normal PRO rings have a puckering amplitude Q between 0.20 and 0.45 Angstrom. If Q is lower than 0.20 Angstrom for a PRO residue, this could indicate disorder between the two different normal ring forms (with C-gamma below and above the ring, respectively). If Q is higher than 0.45 Angstrom something could have gone wrong during the refinement.

  83 PRO  (  86 )     0.10 LOW
 140 PRO  ( 143 )     0.16 LOW

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.

  27 ILE  (  27 )   -2.6171
  21 THR  (  21 )   -2.4165
 121 LYS  ( 124 )   -2.3458
  22 GLU  (  22 )   -2.2220
  14 ARG  (  14 )   -2.1030

Warning: Backbone torsion angle evaluation shows 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.

  52 ASN  (  55 )   Poor phi/psi

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 constrained. This seems to be the case with the current structure, as the observed standard deviation is below 4.0 degrees.

Standard deviation of omega values : 2.402

Note: Ramachandran plot

In this Ramachandran plot X-signs represent glycines, squares represent prolines and small plus-signs represent the other residues. If too many plus-signs fall outside the contoured areas then the molecule is poorly refined (or worse).

In a colour picture, the residues that are part of a helix are shown in blue, strand residues in red. "Allowed" regions for helical residues are drawn in blue, for strand residues in red, and for all other residues in green.

Chain without chain identifier

Accessibility related checks

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 without chain identifier

Bump checks

Error: Abnormally short interatomic distances

The pairs of atoms listed in the table below have an unusually short distance.

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 centers of the two atoms.

The last text-item on each line represents the status of the atom pair. The text `INTRA' means that the bump is between atoms that are explicitly listed in the PDB file. `INTER' means it is an inter-symmetry bump. If the final column contains the text 'HB', the bump criterium 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). If the last column is 'BF', the sum of the B-factors of the atoms is higher than 80, which makes the appearance of the bump somewhat less severe because the atoms probably aren't there anyway.

Bumps between atoms for which the sum of their occupancies is lower than one are not reported. In any case, each bump is listed in only one direction.

 162 CL   ( 173 )     CL     --  174 HOH  ( 291 )              1.117   1.683 INTER
  16 LYS  (  16 )      O     --   17 ILE  (  17 )      C       0.325   2.475 INTRA BF
  25 TYR  (  25 )      N     --   34 THR  (  34 )      CG2     0.310   2.790 INTRA BF
  14 ARG  (  14 )      NH1   --  167 HOH  ( 236 )              0.251   2.299 INTRA BF
  17 ILE  (  17 )      CG2   --   18 TYR  (  18 )      N       0.246   2.854 INTRA
  69 ASP  (  72 )      OD2   --   93 ARG  (  96 )      NH1     0.241   2.309 INTER HB
  16 LYS  (  16 )      CE    --   54 VAL  (  57 )      CG2     0.212   2.988 INTRA BF
  82 LYS  (  85 )      N     --   83 PRO  (  86 )      CD      0.207   2.793 INTRA
  16 LYS  (  16 )      CD    --   54 VAL  (  57 )      CG2     0.207   2.993 INTRA BF
  99 MET  ( 102 )      CE    --  103 MET  ( 106 )      CE      0.200   3.000 INTRA
  11 GLU  (  11 )      OE2   --  142 ARG  ( 145 )      NH1     0.195   2.355 INTRA HB
  26 THR  (  26 )      CG2   --   27 ILE  (  27 )      N       0.167   2.933 INTRA
  20 ASP  (  20 )      O     --   23 GLY  (  23 )      N       0.162   2.388 INTRA BF
 137 ASN  ( 140 )      ND2   --  171 HOH  ( 300 )              0.156   2.394 INTRA HB
  62 LYS  (  65 )      NZ    --  173 HOH  ( 273 )              0.147   2.403 INTER BF
  45 LYS  (  48 )      CE    --  174 HOH  ( 197 )              0.136   2.664 INTER
 142 ARG  ( 145 )      O     --  143 ALA  ( 146 )      C       0.131   2.669 INTRA
 129 ASN  ( 132 )      O     --  130 LEU  ( 133 )      C       0.118   2.682 INTRA
  73 ARG  (  76 )      CD    --  171 HOH  ( 328 )              0.113   2.687 INTER BF
  74 GLY  (  77 )      O     --   75 ILE  (  78 )      C       0.110   2.690 INTRA
  20 ASP  (  20 )      C     --   22 GLU  (  22 )      N       0.109   2.791 INTRA BF
  11 GLU  (  11 )      CD    --  142 ARG  ( 145 )      NH1     0.104   2.996 INTRA
  49 ARG  (  52 )      NH2   --  167 HOH  ( 196 )              0.098   2.452 INTRA BF
  51 THR  (  54 )      C     --   53 GLY  (  56 )      N       0.098   2.802 INTRA BF
   1 MET  (   1 )      CG    --  155 TRP  ( 158 )      CD2     0.094   3.106 INTRA BF
And so on for a total of 65 lines

3D-database related checks

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.

  77 ARG  (  80 )    -5.75
 138 GLN  ( 141 )    -5.53
 151 ARG  ( 154 )    -5.33
 102 GLN  ( 105 )    -5.23
 132 LYS  ( 135 )    -5.17

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 without chain identifier

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 without chain identifier

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 constraints on the neighboring 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!

  32 LEU  (  32 )    3.19   19
  81 LEU  (  84 )    2.75   53
  10 ASP  (  10 )    2.43   51
 151 ARG  ( 154 )    2.26   47
  12 GLY  (  12 )    2.13   11
 138 GLN  ( 141 )    2.04   18
 141 ASN  ( 144 )    1.85   80
  55 ILE  (  58 )    1.55   31

Warning: Unusual backbone conformations

For the residues listed in the table below, the backbone formed by itself and two neighboring 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 center.

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!

  21 THR  (  21 )    0
  29 ILE  (  29 )    0
  47 ILE  (  50 )    0
 103 MET  ( 106 )    0
 111 PHE  ( 114 )    0
 155 TRP  ( 158 )    0
  27 ILE  (  27 )    1
 132 LYS  ( 135 )    1
  51 THR  (  54 )    2
 139 THR  ( 142 )    2

Water related checks

Error: Water clusters without contacts with non-water atoms

The water molecules listed in the table below are part of water molecule clusters that do not make contacts with non-waters. These water molecules are part of clusters that have a distance at least 1 Angstrom larger than the sum of the Van der Waals radii to the nearest non-solvent atom. Because these kinds of water clusters usually are not observed with X-ray diffraction their presence could indicate a refinement artifact. The number in brackets is the identifier of the water molecule in the input file.

 172 HOH  ( 325 )
 172 HOH  ( 333 )
 173 HOH  ( 334 )

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.

 168 HOH  ( 246 )              32.843   17.969  -11.651
 171 HOH  ( 324 )              34.130   -2.577    9.400
 171 HOH  ( 328 )              23.817   -3.141    9.908
 172 HOH  ( 325 )              29.178   24.440   16.767
 172 HOH  ( 326 )              46.775   21.536   33.679
 172 HOH  ( 329 )              43.881   20.882   35.381
 172 HOH  ( 333 )              29.282   18.611    2.894
 173 HOH  ( 296 )              41.751    6.727   26.081
 173 HOH  ( 297 )              40.329    9.145   28.929
 173 HOH  ( 334 )              48.343   -1.238   13.371
 174 HOH  ( 327 )              39.908   23.082    3.638
 174 HOH  ( 332 )              46.431   12.675    5.467

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.

 167 HOH  ( 304 )
 168 HOH  ( 246 )
 169 HOH  ( 227 )
 169 HOH  ( 283 )
 169 HOH  ( 294 )
 169 HOH  ( 295 )
 172 HOH  ( 303 )
 172 HOH  ( 325 )
 172 HOH  ( 326 )
 172 HOH  ( 329 )
 172 HOH  ( 333 )
 173 HOH  ( 250 )
 173 HOH  ( 271 )
 173 HOH  ( 334 )

B-factor analysis

Error: The B-factors of bonded atoms show signs of over-refinement

For each of the bond types in a protein a distribution was derived for the difference between the square roots of the B-factors of the two atoms. All bonds in the current protein were scored against these distributions. The number given below is the RMS Z-score over the structure. For a structure with completely restrained B-factors within residues, this value will be around 0.35, for extremely high resolution structures refined with free isotropic B-factors this number is expected to be near 1.0. Any value over 1.5 is sign of severe over-refinement of B-factors.

RMS Z-score : 3.496 over 1020 bonds
Average difference in B over a bond : 8.86
RMS difference in B over a bond : 13.34

Note: B-factor plot

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

Chain without chain identifier

Hydrogen bond 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 favorable 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. If a residue is marked ``flexible'' the flipped conformation is only slightly better than the non-flipped conformation.

  50 ASN  (  53 )
 137 ASN  ( 140 )

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.

   2 ASN  (   2 )      N
  25 TYR  (  25 )      N
  89 ASP  (  92 )      N

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 constraint values. The first part of the table shows a number of constraint-independent quality indicators.


Structure Z-scores, positive is better than average:

  1st generation packing quality :   0.913
  2nd generation packing quality :   1.482
  Ramachandran plot appearance   :  -0.520
  chi-1/chi-2 rotamer normality  :  -1.413
  Backbone conformation          :  -0.382

RMS Z-scores, should be close to 1.0:
  Bond lengths                   :   0.887
  Bond angles                    :   1.143
  Omega angle restraints         :   0.437 (tight)
  Side chain planarity           :   0.934
  Improper dihedral distribution :   1.065
  B-factor distribution          :   3.496 (loose)
  Inside/Outside distribution    :   0.931

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.

The second part of the table mostly gives an impression of how well the model conforms to common refinement constraint values. The first part of the table shows a number of constraint-independent 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 :   1.4
  2nd generation packing quality :   0.6
  Ramachandran plot appearance   :  -0.2
  chi-1/chi-2 rotamer normality  :  -0.8
  Backbone conformation          :  -0.7

RMS Z-scores, should be close to 1.0:
  Bond lengths                   :   0.887
  Bond angles                    :   1.143
  Omega angle restraints         :   0.437 (tight)
  Side chain planarity           :   0.934
  Improper dihedral distribution :   1.065
  B-factor distribution          :   3.496 (loose)
  Inside/Outside distribution    :   0.931