Profiling protein function with small molecule microarrays.
The regulation of protein function through posttranslational modification, local environment, and protein-protein interaction is critical to cellular function. The ability to analyze on a genome-wide scale protein functional activity rather than changes in protein abundance or structure would provide important new insights into complex biological processes. Herein, we report the application of a spatially addressable small molecule microarray to an activity-based profile of proteases in crude cell lysates. The potential of this small molecule-based profiling technology is demonstrated by the detection of caspase activation upon induction of apoptosis, characterization of the activated caspase, and inhibition of the caspase-executed apoptotic phenotype using the small molecule inhibitor identified in the microarray-based profile.
Winssinger
Nicolas
N
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121, USA.
Ficarro
Scott
S
Schultz
Peter G
PG
Harris
Jennifer L
JL
D000595:Amino Acid Sequence; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D056657:Cathepsin K; D002403:Cathepsins / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000737:chemistry; D020869:Gene Expression Profiling; D006801:Humans; D019169:Jurkat Cells; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D012680:Sensitivity and Specificity
D004791:Enzyme Inhibitors; D011506:Proteins; D002403:Cathepsins; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; C534130:CTSK protein, human; D056657:Cathepsin K