Preparation of the caspase-3/7 substrate Ac-DEVD-pNA by solution-phase peptide synthesis

Quinn P. Peterson, David R. Goode, Diana C. West, Rachel C. Botham, Paul J. Hergenrother

Research output: Contribution to journalArticlepeer-review


This protocol describes the gram-scale solution-phase synthesis of the colorimetric caspase-3/7 substrate Ac-DEVD-pNA. The caspase enzymes are integral to cellular inflammation and apoptotic cascades, and are commonly studied by cell biologists, medicinal chemists and chemical biologists. In particular, the assessment of caspase enzymatic activity is a standard method to evaluate cell death pathways and new apoptosis-modulating agents. Caspase enzymatic activity can be conveniently monitored with peptidic chromogenic or fluorogenic substrates, with certain peptide sequences imparting selectivity for certain caspases. The synthesis of these peptide substrates is typically carried out by solid-phase synthesis, a method that is not ideal for production of the gram quantities needed for high-throughput screening. Described herein is a facile method for the synthesis of the Ac-DEVD-pNA caspase-3/7 substrate using solution-phase peptide synthesis. This protocol, involving iterative PyBOP-mediated couplings and Fmoc deprotections, is rapid (about 5 d), operationally simple and can be used to generate over 1 g of product at a fraction of the cost of the commercial substrate.

Original languageEnglish (US)
Pages (from-to)294-302
Number of pages9
JournalNature Protocols
Issue number2
StatePublished - Feb 2010

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


Dive into the research topics of 'Preparation of the caspase-3/7 substrate Ac-DEVD-pNA by solution-phase peptide synthesis'. Together they form a unique fingerprint.

Cite this