Comprehensive protease specificity profiling

A single-experiment workflow profiles protease cleavage specificity by screening ~1012 octamer peptide permutations, producing specificity maps for ADAM17, streptopain, and factor Xa.

• Why it matters: Many proteases remain poorly characterized, limiting their use in medicine, industry, and research tools; detailed cleavage specificity is needed to guide applications.
• What they did: The approach combines in vitro selection, high-throughput sequencing, mass spectrometry, and automated motif mining, then uses freely available software to convert cleavage data into specificity information.
• The result: Specificity maps corroborate canonical cleavage motifs while extending spectrum preferences, offering insights into broad-specificity proteases like ADAM17 and streptopain.

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