title: Selectivity in ISG15 and ubiquitin recognition by the SARS coronavirus papain-like protease.
abstract: The severe acute respiratory syndrome coronavirus papain-like protease (SARS-CoV PLpro) carries out N-terminal processing of the viral replicase polyprotein, and also exhibits Lys48-linked polyubiquitin chain debranching and ISG15 precursor processing activities in vitro. Here, we used SDS-PAGE and fluorescence-based assays to demonstrate that ISG15 derivatives are the preferred substrates for the deubiquitinating activity of the PLpro. With k(cat)/K(M) of 602,000 M(-1)s(-1), PLpro hydrolyzes ISG15-AMC 30- and 60-fold more efficiently than Ub-AMC and Nedd8-AMC, respectively. Data obtained with truncated ISG15 and hybrid Ub/ISG15 substrates indicate that both the N- and C-terminal Ub-like domains of ISG15 contribute to this preference. The enzyme also displays a preference for debranching Lys48- over Lys63-linked polyubiquitin chains. Our results demonstrate that SARS-CoV PLpro can differentiate between ubiquitin-like modifiers sharing a common C-terminal sequence, and that the debranching activity of the PLpro is linkage type selective. The potential structural basis for the demonstrated specificity of SARS-CoV PLpro is discussed.
authors: Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Avenue, Montreal, Que., Canada H4P 2R2.
authors: Holger A
authors: HA
authors: Lindner
authors: Viktoria
authors: V
authors: Lytvyn
authors: Hongtao
authors: H
authors: Qi
authors: Paule
authors: P
authors: Lachance
authors: Edmund
authors: E
authors: Ziomek
authors: Robert
authors: R
authors: Ménard
mesh_terms: D001665:Binding Sites; D000086782:Coronavirus 3C Proteases; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D016207:Cytokines / Q000737:chemistry*; D004789:Enzyme Activation; D011485:Protein Binding; D013379:Substrate Specificity; D025801:Ubiquitin / Q000737:chemistry*; D014452:Ubiquitins / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D016207:Cytokines; D025801:Ubiquitin; D014452:Ubiquitins; D014764:Viral Proteins; C074034:ISG15 protein, human; D003546:Cysteine Endopeptidases; D000086782:Coronavirus 3C Proteases
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