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Identification of a novel calpain inhibitor using phage display
Journal article   Peer reviewed

Identification of a novel calpain inhibitor using phage display

Rodney P. Guttmann, George A. Day III, Xiaohong Wang and Kara A Bottiggi
Biochemical and Biophysical Research Communications, Vol.333, pp.1087-1092
333
2005
DOI: https://doi.org/10.1016/j.bbrc.2005.06.036
PMID: 15979564
Web of Science ID: WOS:000230538900007

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Abstract

Calpains are calcium- and thiol-dependent proteases that cleave a variety of intracellular substrates. Overactivation of the calpains has been implicated in a number of diseases and conditions such as ischemic stroke indicating a need for the development of calpain inhibitors. A major problem with current calpain inhibitors has been specific targeting to calpain. To identify highly specific calpain interacting peptides, we developed a peptide-phage library screening method based on the calcium-dependent conformation change associated with calpain activation. A phage-peptide library representing greater than 2 billion expressed 12-mers was incubated with calpain I in the presence of calcium. The calcium-dependent bound phage was then eluted by addition of EGTA. After four rounds of selection we found a conserved 5-mer sequence represented by LSEAL. Synthetic LSEAL inhibited tau-calpain interaction and in vitro proteolysis of tau- and alpha-synuclein by calpains. Deletion of the portion of the tau protein containing a homologous sequence to LSEAL resulted in decreased calpain-mediated tau degradation. These data suggest that these peptides may represent novel calpastatin mimetics.

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