Description - Procedure - Analysis - Complete worked example (Zip file, 242 Kb)
When COMPARE is present, a pair of protein systems is compared to identify similarities and differences in their geometries. These systems can be PDB or ENT files, MOPAC data sets, i.e., files that end in ".mop" or ".dat", or MOPAC archive files, i.e., files that end in ".arc". The comparison is done using a MOPAC data-set that contains information on the locations of the systems to be used. A useful practice is to have the systems in one or two folders and the data-set for the comparison in another folder at the same level, e.g., if the systems are in "Users/name/Crambin/Original PDB files" and "Users/name/Crambin/MOPAC files" then the compare files should be in "/Users/name/Crambin/Compare"
In a normal run, atoms of one system are re-arranged to match those of the other system, so that both systems have the same atoms in the same sequence. The geometry of one system, "System A", is then rotated and translated to minimize the difference between it and that of "System B." Once the systems are in maximum coincidence, an analysis is performed; this identifies differences in the systems, differences in bond-lengths, and other quantities, such as the Root-Mean-Square and average atomic position differences.
In the PDB format there are two systems for labeling hydrogen atoms. MOPAC uses the older system. If one file uses the new system and the other uses the old system, the file that uses the new system can be converted into the old one by keyword SITE=().
If LET is present, then hydrogen atoms that are equivalent in the two systems, but have different names, will be treated as if they were equivalent. Normally, these atoms would be excluded from the analysis, but it's useful to include them when different steps in a reaction mechanism are being compared. This is particularly useful when hydrogen atoms (protons or hydride ions) migrate from one atom to another, for example when a hydroxyl group ionizes.
The recommended way to use COMPARE is to use a data set, e.g. "compare the geometries of system-A and system-B.mop", that contains three lines of the type:
COMPARE geo_dat="System A" geo_ref="System B" output Text to be displayed Second line of text to be displayed (Optional)In this form, with OUTPUT present, the amount of output is minimized and only data related to the comparison is printed. A web-page in HTML, e.g., "compare the geometries of system-A and system-B.html", is written that allows a very rapid visual comparison of features.
The text "System A" and "System B" should be replaced with the file-names for the systems of interest. For example, to compare a protein system "1A1A PDB structure.mop" and "1A1A optimized.arc" the first line would consist of:
COMPARE geo_dat="1A1A optimized.arc" geo_ref="1A1A PDB structure.mop" output Compare original PDB structure and optimized geometry for system 1A1A
NOSWAP: By default, atoms can be swapped in the GEO_REF data-set if that would increase the overlap with the GEO_DAT data set. To prevent this swapping, add NOSWAP.
NOREOR: Do not move the GEO_REF system to minimize the difference between it and the GEO_DAT system.
OUTPUT: Not essential, but reduced the size of the output, and allows the output of interest to be located more rapidly.
Optional, but useful. These lines will be displayed in the HTML web-page, and an informative description of the system can be very helpful later on when several web-pages are being used.
"Differences in atoms sets" - Self-explanatory. Have a quick look at the differences, and if nothing unexplained is seen, ignore this section.
"Number of atoms" - Self-explanatory.
"After docking - Atoms that move a lot" - This list is in order of decreasing motion. The first entry is the atom that shows the largest motion in going from one system to the other. A useful operation would be to open the HTML web-page generated by the run, and look at some of the atoms that move a lot.
"Residues that move a lot and their (Movement in Angstroms per atom)" - This list is in order of decreasing motion. The first entry is the residue that shows the largest motion in going from one system to the other.
In the next few entries, the assumption is made that one geometry is a reference, e.g., a PDB file downloaded from the internet, and the other is a geometry optimized using MOPAC.
"Difference between GEO_DAT and GEO_REF:"
"total"
is the sum of the differences in atom positions.
"Average" is "total"
divided by the number of atoms.
"RMS, in Angstroms" is the
root-mean-square of the differences between the two geometries. This is strictly comparable with the RMSD quantities reported
in other fields of protein structural chemistry, e.g., different PDB reports on
the same system.
In the HTML file, "Diff." is the same as the "total" in the output file, and "RMSD" is the same as "RMS" in the output file.
"Differences between bond-lengths for the two geometries" - This is important. There are three common reasons for large (greater than 0.05 or 4%) differences in bond-length: (A) faults in the PDB structure, (B) faults in the computational method being used, e.g., PM7 with COSMO, and (C) incorrect hydrogenation resulting in a local fault in the chemical structure. To identify where the fault lies, examine the bond using the HTML web-page. In order to get a better view of the fault, display only the local environment of the two atoms. If the fault is identified as (A), ignore the fault - it was corrected by the geometry optimization. If (B), also ignore the fault, but make a note that this is a fault in the method being used, and might be important. If you think it is important, try using a different method, e.g., PM6-D3H4 with COSMO.
Most PDB structures do not include hydrogen atoms, and do not distinguish between the carboxylic acid C-O distances or between arginine CZ and (NH1 or NH2) distances. When a geometry is optimized, pairs of C-O and C-N bonds that PDB reports are the same length will be predicted to have distinctly different lengths, one will be a single bond and the other a double bond. This causes a difference in bond-lengths to be reported, but these differences are not a result of a fault in either the PDB or the semiempirical structure. Instead, these differences should be ignored. For example, the following Table lists the pairs of atoms that show the largest differences in bond-lengths. All the atom-pairs that involve carboxylate atoms, e.g. GLU CD and (OE1 or OE2), ASP CG and (OD1 or OD2) or guanidine atoms in ARG CZ and (NH1 or NH2) should be ignored.
Both PM6 and PM7 over-estimate the peptide C-N bond-length by about 0.08 Å, at about 1.37 instead of the expected 1.29 Å.
As a worked example, look at the Table
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Table showing differences in bond-lengths between PDB and calculated geometries |
Differences between bond-lengths for the two geometries
Diff. Atom 1 Atom 2 Bond length Bond length
in GEO_DAT in GEO_REF 1 0.148 O(ATOM 524 OE2 GLU A 37) C(ATOM 522 CD GLU A 37) 1.368 1.219 2 0.139 O(ATOM 1086 OD2 ASP A 74) C(ATOM 1084 CG ASP A 74) 1.348 1.209 3 0.112 N(ATOM 3864 NH1 ARG A 258) C(ATOM 3863 CZ ARG A 258) 1.436 1.324 4 0.111 O(ATOM 3432 OE2 GLU A 231) C(ATOM 3430 CD GLU A 231) 1.349 1.238 5 0.109 N(ATOM 1513 NH1 ARG A 100) C(ATOM 1512 CZ ARG A 100) 1.423 1.314 6 0.102 N(ATOM 3930 NE ARG A 262) C(ATOM 3931 CZ ARG A 262) 1.438 1.336 7 0.100 N(ATOM 3175 NE ARG A 216) C(ATOM 3176 CZ ARG A 216) 1.413 1.313 8 0.097 O(ATOM 523 OE1 GLU A 37) C(ATOM 522 CD GLU A 37) 1.216 1.314 9 0.096 N(ATOM 932 N GLN A 64) C(ATOM 913 C TYR A 63) 1.391 1.295 10 0.094 N(ATOM 1511 NE ARG A 100) C(ATOM 1512 CZ ARG A 100) 1.413 1.319 11 0.092 N(ATOM 1961 NH1 ARG A 132) C(ATOM 1960 CZ ARG A 132) 1.422 1.330 12 0.091 N(ATOM 315 N ASN A 23) C(ATOM 307 C ALA A 22) 1.385 1.294 13 0.090 N(ATOM 1355 NH1 ARG A 91) C(ATOM 1354 CZ ARG A 91) 1.410 1.320 14 0.089 N(ATOM 3729 N THR A 250) C(ATOM 3711 C PHE A 249) 1.378 1.288 15 0.087 N(ATOM 1016 N GLU A 70) C(ATOM 1011 C GLY A 69) 1.400 1.313 16 0.086 N(ATOM 3177 NH1 ARG A 216) C(ATOM 3176 CZ ARG A 216) 1.420 1.335 17 0.086 N(ATOM 3125 N THR A 213) C(ATOM 3108 C ILE A 212) 1.379 1.293 18 0.081 N(ATOM 3841 N THR A 257) C(ATOM 3832 C SER A 256) 1.406 1.325 19 0.081 O(ATOM 3431 OE1 GLU A 231) C(ATOM 3430 CD GLU A 231) 1.227 1.308 20 0.081 N(ATOM 580 N THR A 42) C(ATOM 575 C GLY A 41) 1.379 1.298 21 0.080 N(ATOM 3492 N GLU A 235) C(ATOM 3480 C PRO A 234) 1.378 1.298 22 0.080 N(ATOM 1959 NE ARG A 132) C(ATOM 1960 CZ ARG A 132) 1.417 1.337 23 0.080 N(ATOM 305 N ALA A 22) C(ATOM 300 C GLY A 21) 1.390 1.310 24 0.080 N(ATOM 679 N VAL A 48) C(ATOM 664 C HIS A 47) 1.372 1.292 25 0.080 N(ATOM 1822 NE2 HIS A 122) C(ATOM 1823 CE1 HIS A 122) 1.404 1.324 |
IMPORTANT: If the fault is caused by incorrect hydrogenation, than edit the optimized geometry to correct the fault, and re-run the geometry optimization. Then re-run the comparison to verify that the fault has been corrected. Incorrect hydrogenation most often occurs in substrates and hetero-groups such as heme rings. A lot of grief can be avoided by looking at differences in bond-lengths and checking for hydrogenation errors. For examples of PDB files that have severe errors in bond-lengths, see Preparing a starting data set.
Other global data such as the date, heats of formation, number of atoms, etc., are printed in the output and in the HTML web-page.