Use of protease proteomics to discover granzyme B substrates

Andrew J. Bredemeyer, R. Reid Townsend, Timothy J. Ley

Research output: Contribution to journalReview articlepeer-review

12 Scopus citations

Abstract

A wide variety of proteases play important roles in immunity, including the destruction of microbes, induction of apoptosis, antigen processing, and regulation of the immune response. Characterization of these proteases requires not only an understanding of substrate specificity, but also the identification of specific protein substrates. Recent advances in proteomics technology have introduced new techniques for the study of protease function. Here, we highlight a proteomic approach used in our laboratory that employs two-dimensional gel electrophoresis coupled with mass spectrometry to identify native protease substrates. With this technique, we have successfully detected both known and novel granzyme B substrates, characterized cleavage products, and identified a granzyme B cleavage site. This approach may serve as an important discovery tool for other immunologic proteases.

Original languageEnglish
Pages (from-to)143-153
Number of pages11
JournalImmunologic Research
Volume32
Issue number1-3
DOIs
StatePublished - 2005

Keywords

  • Caspases
  • Granzyme B
  • Proteases
  • Proteomics
  • Two-dimensional difference in-gel electrophoresis

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