Can I read A Broad‐spectrum Fluorescence‐based Peptide Library for the Rapid Identification of Protease Substrates on EtoBox?

A Broad‐spectrum Fluorescence‐based Peptide Library for the Rapid Identification of Protease Substrates by Daniel A. Thomas; Peter Francis; Carla Smith; Steven Ratcliffe; Nicholas J. Ede; Corinne Kay; Gareth Wayne; Steve L. Martin; Keith Moore; Augustin Amour; Nigel M. Hooper is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is A Broad‐spectrum Fluorescence‐based Peptide Library for the Rapid Identification of Protease Substrates about?

## Abstract Identification of peptide substrates for proteases can be a major undertaking. To overcome issues such as feasibility and deconvolution, associated with large peptide libraries, a ‘small but smart’ generic fluorescence resonance energy transfer rapid endopeptidase profiling library (REPLi) was synthesised as a tool for rapidly identifying protease substrates. Within a tripeptide core, flanked by Gly residues, similar amino acids were paired giving rise to a relatively small library of 3375 peptides divided into 512 distinct pools each containing only 8 peptides. The REPLi was validated with trypsin, pepsin, the matrix metalloprotease (MMP)‐12 and MMP‐13 and calpains‐1 and ‐2. In the case of calpain‐2, a single iteration step involving LC‐MS, provided the definitive residue specificity from which a highly sensitive fluorogenic substrate, (FAM)‐Gly‐Gly‐Gly‐Gln‐Leu‐Tyr‐Gly‐Gly‐DPA‐Arg‐Arg‐Lys‐(TAMRA), was then designed. The thorough validation of this ‘small but smart’ peptide library with representatives from each of the four mechanistic protease classes indicates that the REPLi will be useful for the rapid identification of substrates for multiple proteases.

Who reads A Broad‐spectrum Fluorescence‐based Peptide Library for the Rapid Identification of Protease Substrates?

Author

Daniel A. Thomas; Peter Francis; Carla Smith; Steven Ratcliffe; Nicholas J. Ede; Corinne Kay; Gareth Wayne; Steve L. Martin; Keith Moore; Augustin Amour; Nigel M. Hooper

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley; Research Square (ISSN 1615-9853)

Published

2006

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)