An innovative application of the "flexible" GRID/PCA computational method: study of differences in selectivity between PGAs from Escherichia coli and a Providentia rettgeri mutant.
The original GRID/PCA technique was adapted for the development of a tool potentially useful for the plan of a research strategy in rational enzyme design. The use of the MOVE directive of GRID made it possible to partially take into account protein flexibility, and the multivariate analysis was used as an instrument for focusing only on relevant information related to the differences in enzyme substrate selectivities. The comparison of two different penicillin G acylases, from Escherichia coli and from Providentia rettgeri, was used as a case study; these enzymes are very similar and their reported selectivities differ only for a couple of mutations around the active site. The "flexible" GRID/PCA method was able to correctly predict the observed selectivity differences caused not only by mutations of residues of the active site but also by long range effects on substrate selectivity due to sequence mutations on residues not directly involved in substrate recognition.
Dipartimento di Scienze Farmaceutiche, Università degli Studi, piazzale Europa 1, 34127 Trieste, Italy.
Paolo
P
Braiuca
Gabriele
G
Cruciani
Cynthia
C
Ebert
Lucia
L
Gardossi
Paolo
P
Linda
D001419:Bacteria / Q000201:enzymology*; D019295:Computational Biology; D004926:Escherichia coli / Q000201:enzymology*; D015335:Molecular Probes; D009154:Mutation; D010399:Penicillin Amidase / Q000378:metabolism*; D013379:Substrate Specificity
D015335:Molecular Probes; D010399:Penicillin Amidase