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Deubiquitylase, deSUMOylase, and deISGylase activity microarrays for assay of substrate preference and functional modifiers.

Authors
  • Loch, Christian M
  • Cuccherini, Charles L
  • Leach, Craig A
  • Strickler, James E
Type
Published Article
Journal
Molecular & Cellular Proteomics
Publisher
American Society for Biochemistry and Molecular Biology
Publication Date
Jan 01, 2011
Volume
10
Issue
1
Identifiers
DOI: 10.1074/mcp.M110.002402
PMID: 20956615
Source
Medline
License
Unknown

Abstract

Microarray-based proteomics expanded the information potential of DNA arrays to the level of protein translation and interaction, but so far, not much beyond. Although enzymatic activity from immobilized proteins has been reliably studied using surface plasmon resonance, a microarray of catalytically competent enzymes would facilitate high throughput, parallel study of their function. The ability to localize activity from soluble substrates has frustrated development of such an array. Here, we report the novel use of previously developed, highly specific suicide substrates for three families of enzymes: deubiquitylases, deSUMOylases, and deISGylases. We show specificity of each family to its cognate substrate, and demonstrate utility of the array in a secondary screen of small molecule inhibitors.

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