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. Author manuscript; available in PMC: 2013 Oct 24.

Published in final edited form as: J Am Chem Soc. 2012 Oct 11;134(42):17704–17713. doi: 10.1021/ja307599z

Design of a calpain specific ABP (top). ABPs contain a mechanism based inhibitor, specificity element, and tag. Only the chemical structures ABPs containing a biotin tag are shown here. denotes the m-xylyl c15 crosslinking between the cysteines. ABP binding to calpain-1 (bottom). The linker length and rigidity between the crosslinked peptide and succinyl epoxide was evaluated via reaction with calpain-1 in vitro. A five-carbon backbone, flexible linker appears optimal. Loading control lanes beneath the panel show Western blot analysis using anti-calpain-1.

Inline graphic — Design of a calpain specific ABP (top). ABPs contain a mechanism based inhibitor, specificity element, and tag. Only the chemical structures ABPs containing a biotin tag are shown here. denotes the m-xylyl c15 crosslinking between the cysteines. ABP binding to calpain-1 (bottom). The linker length and rigidity between the crosslinked peptide and succinyl epoxide was evaluated via reaction with calpain-1 in vitro. A five-carbon backbone, flexible linker appears optimal. Loading control lanes beneath the panel show Western blot analysis using anti-calpain-1.