A series of penicillin-derived C2-symmetric inhibitors of HIV-1 proteinase: structural and modeling studies.

The binding modes of a series of penicillin-derived C2 symmetric dimer inhibitors of HIV-1 proteinase were investigated by NMR, protein crystallography, and molecular modeling. The compounds were found to bind in a symmetrical fashion, tracing and S-shaped course through the active site, with good hydrophobic interactions in the S1/S1' and S2/S2' pockets and hydrogen bonding of inhibitor amide groups. Interactions with the catalytic aspartates appeared poor and the protein conformation was very similar to that seen in complexes with peptidomimetics, in spite of the major differences in ligand structure.

Knowledge based modelling of homologous proteins, Part II: Rules for the conformations of substituted sidechains.

This paper describes a rapid, automated procedure which can be used for model building sidechains using (i) spatial information from sidechains in topologically equivalent positions as far as such a correlation is observed, and then (ii) most probable conformations of the sidechains in the respective secondary structure type. Analysis of topologically equivalent residues in the structurally conserved regions of a family of proteins implies that the spatial positions of the atoms in the sidechains rather than conformations should be considered when model building. Rules for the modelling of all 20 side-chains from each other in alpha-helical, beta-sheet and loop regions--a total of 1200--are established. Cluster analysis is used on positional data from the sidechain atoms of structurally equivalent residues in an homologous family to guide modelling. The most probable conformation for the sidechain is used for modelling atoms where no useful guidance is obtainable from equivalent sidechains of the homologous proteins. In order to test the procedure we have modelled the sidechains of the residues in the structurally conserved regions of myoglobin from four other globins. The automated procedure described here has been incorporated into the program COMPOSER.

Prediction of electrostatic effects of engineering of protein charges.

Accurate prediction of electrostatic effects on catalytic activity is an essential component of protein design. Site-directed mutagenesis of charged groups in subtilisin of Bacillus amyloliquefaciens has provided experimental measurements of electrostatic interactions which may be used to test such theoretical methods. The pKa of the histidine of the active site has been perturbed by +0.08 to -1.0 units by modifying one or two residues. Electrostatic effects in proteins can be modelled by the algorithm of Warwicker and Watson, which uses classical electrostatics and considers both the charge position and the shape of the molecule. Here we report that the algorithm can model several pKa shifts in subtilisin to fair accuracy.