1. Substrate specificity of Staphylococcus aureus cysteine proteases – Staphopains A, B and C
- Author
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Kalińska, Magdalena, Kantyka, Tomasz, Greenbaum, Doron C., Larsen, Katrine S., Władyka, Benedykt, Jabaiah, Abeer, Bogyo, Matthew, Daugherty, Patrick S., Wysocka, Magdalena, Jaros, Marcelina, Lesner, Adam, Rolka, Krzysztof, Schaschke, Norbert, Stennicke, Henning, Dubin, Adam, Potempa, Jan, and Dubin, Grzegorz
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STAPHYLOCOCCUS aureus , *CYSTEINE proteinases , *FLUORESCENCE resonance energy transfer , *NITROBENZOIC acid , *GREEN fluorescent protein , *IMMUNOSPECIFICITY - Abstract
Abstract: Human strains of Staphylococcus aureus secrete two papain-like proteases, staphopain A and B. Avian strains produce another homologous enzyme, staphopain C. Animal studies suggest that staphopains B and C contribute to bacterial virulence, in contrast to staphopain A, which seems to have a virulence unrelated function. Here we present a detailed study of substrate preferences of all three proteases. The specificity of staphopain A, B and C substrate-binding subsites was mapped using different synthetic substrate libraries, inhibitor libraries and a protein substrate combinatorial library. The analysis demonstrated that the most efficiently hydrolyzed sites, using Schechter and Berger nomenclature, comprise a P2–Gly↓Ala(Ser) sequence motif, where P2 distinguishes the specificity of staphopain A (Leu) from that of both staphopains B and C (Phe/Tyr). However, we show that at the same time the overall specificity of staphopains is relaxed, insofar as multiple substrates that diverge from the sequences described above are also efficiently hydrolyzed. [Copyright &y& Elsevier]
- Published
- 2012
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