Deubiquitylase, DeSUMOylase, and DeISGylase Activity Microarrays for Assay of Substrate Preference and Functional Modifiers

作者:Loch Christian M*; Cuccherini Charles L; Leach Craig A; Strickler James E
来源:Molecular & Cellular Proteomics, 2011, 10(1).
DOI:10.1074/mcp.M110.002402

摘要

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. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.002402, 1-11, 2011.

  • 出版日期2011-1