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A strategy to profile prime and non-prime proteolytic substrate specificity.

Petrassi, Michael, Williams, Jennifer, Li, Jun, Tumanut, Christine, Ek, Jared, Nakai, Takashi, Masick, Brian, Backes, Bradley and Harris, Jennifer (2005) A strategy to profile prime and non-prime proteolytic substrate specificity. Bioorganic & Medicinal Chemistry Letters, 15 (12). pp. 3162-3166. ISSN 0960-894X


A strategy was developed to determine the prime and non-prime substrate specificity of serine, threonine and cysteine proteases. ACC positional scanning technology was employed to determine the P4-P1 non-prime site substrate specificity. The data was used to synthesize biased donor-quencher positional scanning libraries to profile the P1'-P4' prime site substrate specificity. Directed sorting using the Irori Nanokan system allowed for the archiving of multiple P1'-P4' positional scanning libraries. From these libraries focused donor-quencher libraries incorporating P4-P1 data for each protease under study could be rapidly prepared. The profiling of thrombin and caspase-3 P4-P4' substrate specificity, comparison of the library specificity data to single substrates, and the analysis of physiological cleavage sites are described.

Item Type: Article
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Additional Information: author can archive post-print (ie final draft post-refereeing); Publisher's version/PDF cannot be used
Keywords: Protease substrate specificity; Positional scanning library; Donor–quencher substrate; Directed sorting
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Date Deposited: 14 Dec 2009 14:02
Last Modified: 31 Jan 2013 01:22


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