ORGN 487 |
Chemoselective ligation and activity-based protein profiling assisted by click chemistry
| Lifu Ma1, Anna Katrin Szardenings1, Benjamin F. Cravatt2, and Erik J. Sorensen2. (1) Chemistry, ActivX Biosciences, Inc, 11025 N. Torrey Pines Road, La Jolla, CA 92037, (2) Departments of Chemistry and Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 |
Chemoselective labeling of proteins in a proteomic mixture by activity-based probes is a crucial step for the subsequent analysis. Herein we present an improved bio-conjugation method via the cycloaddition reaction of an azide with an alkyne to form a chemically-stable triazole ring in aqueous solution (“click chemistry”). This approach consists of a two-step in situ process: labeling of proteins with an activity-based probe terminating in an alkyne followed by the introduction of a fluorescent or biotin tag through Cu(I) catalyzed click chemistry. Our improved procedure results in lower background and excellent protein profiling results as demonstrated by gel and LC/MS/MS analysis. Since the conditions are “bio-orthogonal” with protein residues and other functional groups, this procedure represents an alternative approach for protein modification and bio-conjugation applications. |
| |
|
Total Synthesis, Asymmetric Reactions and Syntheses, Bioorganic
9:00 AM-11:00 AM, Wednesday, March 31, 2004 Anaheim Convention Center -- Hall C, Poster
Sci-Mix
8:00 PM-10:00 PM, Monday, March 29, 2004 Anaheim Convention Center -- Hall A, Sci-Mix
Division of Organic Chemistry
The 227th ACS National Meeting, Anaheim, CA, March 28-April 1, 2004
|