title: Identification of the proteasome inhibitor MG262 as a potent ATP-dependent inhibitor of the Salmonella enterica serovar Typhimurium Lon protease.
abstract: Lon is a homo-oligomeric ATP-dependent serine protease which functions in the degradation of damaged and certain regulatory proteins. The importance of Lon activity in bacterial pathogenicity has led to its emergence as a target in the development of novel antibiotics. As no potent inhibitors of Lon activity have been reported to date, we sought to identify an inhibitor which could serve as a lead compound in the development of a potent Lon-specific inhibitor. To determine whether a nucleotide- or peptide-based inhibitor would be more effective, we evaluated the steady-state kinetic parameters associated with both ATP and peptide hydrolysis by human and Salmonella enterica serovar Typhimurium Lon. Although the ATP hydrolysis activities of both homologues are kinetically indistinguishable, they display marked differences in peptide substrate specificity. This suggests that a peptide-based inhibitor could be developed which would target bacterial Lon, thereby decreasing side-effects due to cross-reactivity with human Lon. Using Salmonella enterica serovar Typhimurium Lon as a model, we evaluated the IC50 values of a series of commercially available peptide-based inhibitors. Those inhibitors which behave as transition state analogues were the most useful in inhibiting Lon activity. The peptidyl boronate, MG262, was the most potent inhibitor tested (IC50 = 122 +/- 9 nM) and required binding, but not hydrolysis, of ATP to initiate inhibition. We hope to use MG262 as a lead compound in the development of future Lon-specific inhibitors.
authors.affiliation: Department of Chemistry, Case Western Reserve University, Cleveland, Ohio 44106, USA.
authors.forename: Hilary
authors.initials: H
authors.lastname: Frase
authors.forename: Jason
authors.initials: J
authors.lastname: Hudak
authors.forename: Irene
authors.initials: I
authors.lastname: Lee
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D001897:Boronic Acids / Q000737:chemistry / Q000494:pharmacology*; D003001:Cloning, Molecular; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D015195:Drug Design; D006801:Humans; D020128:Inhibitory Concentration 50; D049070:Protease La / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D061988:Proteasome Inhibitors; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D019779:Salmonella enterica / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D001897:Boronic Acids; D015853:Cysteine Proteinase Inhibitors; C423917:MG 262; D061988:Proteasome Inhibitors; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D049070:Protease La