Citation

Loch CM, Cuccherini CL, Leach CA, Strickler JE. Mol. Cell Proteomics 2011; 10(1): M110.002402.

Copyright

(Copyright © 2011, American Society for Biochemistry and Molecular Biology)

DOI

10.1074/mcp.M110.002402

PMID

20956615

PMCID

PMC3013450

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.

Language: en

Keywords

Humans; Reproducibility of Results; Substrate Specificity; Enzyme Inhibitors; Enzyme Assays; Esters; Enzymes, Immobilized; Endopeptidases; Protein Array Analysis; Small Molecule Libraries; Small Ubiquitin-Related Modifier Proteins; Staining and Labeling; Ubiquitins