Structure and protective efficacy of the Staphylococcus aureus autocleaving protease EpiP. Issue 4 (13th January 2014)

• Record Type: Journal Article
• Authors: Kuhn, Misty L.; Prachi, Prachi; Minasov, George; Shuvalova, Ludmilla; Ruan, Jiapeng; Dubrovska, Ievgeniia; Winsor, James; Giraldi, Monica; Biagini, Massimiliano; Liberatori, Sabrina; Savino, Silvana; Bagnoli, Fabio; Anderson, Wayne F.; Grandi, Guido
• Is Part Of: FASEB journal. Volume 28:Issue 4(2014)
• Page Start: 1780
• Page End: 1793
• Publication Date: 2014-01-13
• Subjects: vaccine -- epidermin leader peptide processing serine protease -- pathogenesis -- lantibiotic -- abscess -- immunogenicity; Biology -- Periodicals; Biology, Experimental -- Periodicals; 570
• DOI: 10.1096/fj.13-241737
• Languages: English
• ISSNs: 0892-6638

Abstract

Abstract : Despite the global medical needs associated with Staphylococcus aureus infections, no licensed vaccines are currently available. We identified and characterized a protein annotated as an epidermin leader peptide processing serine protease (EpiP), as a novel S. aureus vaccine candidate. In addition, we determined the structure of the recombinant protein (rEpiP) by X‐ray crystallography. The crystal structure revealed that rEpiP was cleaved somewhere between residues 95 and 100, and we found that the cleavage occurs through an autocatalytic intramolecular mechanism. The protein expressed by S. aureus cells also appeared to undergo a similar processing event. To determine whether the protein acts as a serine protease, we mutated the hypothesized catalytic serine 393 residue to alanine, generating rEpiP‐S393A. The crystal structure of this mutant protein showed that the polypeptide chain was not cleaved and was not interacting stably with the active site. Indeed, rEpiP‐S393A was shown to be impaired in its protease activity. Mice vaccinated with rEpiP were protected from S. aureus infection (34% survival, P = 0.0054). Moreover, the protective efficacy generated by rEpiP and rEpiP‐S393A was comparable, implying that the noncleaving mutant could be used for vaccination purposes.—Kuhn, M. L., Prachi, P., Minasov, G., Shuvalova, L., Ruan, J., Dubrovska, I., Winsor, J., Giraldi, M., Biagini, M., Liberatori, S., Savino, S., Bagnoli, F., Anderson, W. F., Grandi, G. Structure and protective efficacy of the Staphylococcus aureus autocleaving protease EpiP. FASEB J. 28, 1780–1793 (2014). www.fasebj.org