Phosphoproteomics: Miles To Go Before It's Routine

摘要

Researchers see major technological advances but still have significant challenges to overcome. Not so long ago, the process for determining protein phosphorylation was a labor-intensive and serial process. Typically, a researcher would use ~(32)P-labeling coupled with 2D gel electrophoresis phosphopeptide mapping, autoradiography, band excision, and Edman sequencing. In addition to expending much time and effort, researchers had to work with radioactivity, and the method could not be multiplexed. Now in 2009, two complementary techniques for identification of phosphorylation sites, MS and microarrays, have provided significant increases in throughput. In discovery mode, researchers can identify thousands of phosphorylation sites with LC/MS~(n), and in profiling mode, researchers can detect thousands of phosphorylation events and predict the substrates of various protein kinases with planar, suspension, and reverse microarrays.