PNA-encoded protease substrate microarrays

Chem Biol. 2004 Oct;11(10):1351-60. doi: 10.1016/j.chembiol.2004.07.015.

Abstract

Our current understanding of the role and regulation of protease activity in normal and pathogenic processes is limited by our ability to measure and deconvolute their enzymatic activity. To address this limitation, an approach was developed that utilizes rhodamine-based fluorogenic substrates encoded with PNA tags. The PNA tags address each of the substrates to a predefined location on an oligonucleotide microarray through hybridization, thus allowing the deconvolution of multiple signals from a solution. A library of 192 protease substrates was prepared by split and mix combinatorial synthesis. The methodology and validation of this approach for profiling proteolytic activity from single proteases and from those in crude cell lysates as well as clinical blood samples is described.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Hydrolysis
  • Jurkat Cells
  • Oligonucleotide Array Sequence Analysis / methods*
  • Peptide Hydrolases / chemistry
  • Peptide Hydrolases / genetics*
  • Peptide Hydrolases / metabolism*
  • Peptide Library
  • Peptide Nucleic Acids / chemistry
  • Peptide Nucleic Acids / genetics*
  • Peptide Nucleic Acids / metabolism
  • Proteomics / methods
  • Substrate Specificity / genetics

Substances

  • Peptide Library
  • Peptide Nucleic Acids
  • Peptide Hydrolases