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

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 catalyticall...

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Bibliographic Details
Published in:	Molecular & cellular proteomics Vol. 10; no. 1; p. M110.002402
Main Authors:	Loch, Christian M., Cuccherini, Charles L., Leach, Craig A., Strickler, James E.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-01-2011 The American Society for Biochemistry and Molecular Biology
Subjects:	Endopeptidases - metabolism Enzyme Assays Enzyme Inhibitors - pharmacology Enzymes, Immobilized - metabolism Esters - metabolism Humans Protein Array Analysis - methods Reproducibility of Results Small Molecule Libraries - pharmacology Small Ubiquitin-Related Modifier Proteins - antagonists & inhibitors Small Ubiquitin-Related Modifier Proteins - metabolism Staining and Labeling Substrate Specificity - drug effects Ubiquitins - metabolism
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	1535-9476 1535-9484
DOI:	10.1074/mcp.M110.002402