title: Comparison of the binding specificity of two bacterial metalloproteases, LasB of Pseudomonas aeruginosa and ZapA of Proteus mirabilis, using N-alpha mercaptoamide template-based inhibitor analogues.
abstract: The metalloproteases ZapA of Proteus mirabilis and LasB of Pseudomonas aeruginosa are known to be virulence factors their respective opportunistic bacterial pathogens, and are members of the structurally related serralysin and thermolysin families of bacterial metalloproteases respectively. Secreted at the site of infection, these proteases play a key role in the infection process, contributing to tissue destruction and processing of components of the host immune system. Inhibition of these virulence factors may therefore represent an antimicrobial strategy, attenuating the virulence of the infecting pathogen. Previously we have screened a library of N-alpha mercaptoamide dipeptide inhibitors against both ZapA and LasB, with the aim of mapping the S1' binding site of the enzymes, revealing both striking similarities and important differences in their binding preferences. Here we report the design, synthesis, and screening of several inhibitor analogues, based on two parent inhibitors from the original library. The results have allowed for further characterization of the ZapA and LasB active site binding pockets, and have highlighted the possibility for development of broad-spectrum bacterial protease inhibitors, effective against enzymes of the thermolysin and serralysin metalloprotease families.
authors.affiliation: School of Pharmacy, Queens University Belfast, Medical Biology Centre, 97 Lisburn Road, Belfast BT9 7BL, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Carson
authors.forename: George R
authors.initials: GR
authors.lastname: Cathcart
authors.forename: Howard
authors.initials: H
authors.lastname: Ceri
authors.forename: Brian
authors.initials: B
authors.lastname: Walker
authors.forename: Brendan F
authors.initials: BF
authors.lastname: Gilmore
mesh_terms: D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry*; D004151:Dipeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D008666:Metalloendopeptidases / Q000737:chemistry*; D045726:Metalloproteases / Q000737:chemistry*; D019151:Peptide Library; D011480:Protease Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D011485:Protein Binding; D011513:Proteus mirabilis / Q000187:drug effects / Q000201:enzymology / Q000472:pathogenicity; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000201:enzymology / Q000472:pathogenicity; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004151:Dipeptides; C406102:N-mercaptoamide; D019151:Peptide Library; D011480:Protease Inhibitors; D013438:Sulfhydryl Compounds; D045726:Metalloproteases; D008666:Metalloendopeptidases; C043859:pseudolysin, Pseudomonas aeruginosa