title: Substrate specificity of Staphylococcus aureus cysteine proteases--Staphopains A, B and C.
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.
authors.affiliation: Department of Microbiology, Faculty of Biochemistry Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kalińska
authors.forename: Tomasz
authors.initials: T
authors.lastname: Kantyka
authors.forename: Doron C
authors.initials: DC
authors.lastname: Greenbaum
authors.forename: Katrine S
authors.initials: KS
authors.lastname: Larsen
authors.forename: Benedykt
authors.initials: B
authors.lastname: Władyka
authors.forename: Abeer
authors.initials: A
authors.lastname: Jabaiah
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Daugherty
authors.forename: Magdalena
authors.initials: M
authors.lastname: Wysocka
authors.forename: Marcelina
authors.initials: M
authors.lastname: Jaros
authors.forename: Adam
authors.initials: A
authors.lastname: Lesner
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Rolka
authors.forename: Norbert
authors.initials: N
authors.lastname: Schaschke
authors.forename: Henning
authors.initials: H
authors.lastname: Stennicke
authors.forename: Adam
authors.initials: A
authors.lastname: Dubin
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Dubin
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001667:Binding, Competitive; D001717:Birds; D020134:Catalytic Domain; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D057056:Cysteine Proteases / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D011485:Protein Binding; D054852:Small Molecule Libraries; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000201:enzymology / Q000472:pathogenicity; D013379:Substrate Specificity; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D054852:Small Molecule Libraries; D057056:Cysteine Proteases; D003546:Cysteine Endopeptidases; C501295:staphopain A protein, Staphylococcus aureus; C479137:staphopain B protein, Staphylococcus aureus