Determination of the complex secretory proteome of adipocytes and its metabolic changes induced by drug treatment such as insulin or rosiglitazone is possible with the advanced proteomics technologies described herein. To study the secreted proteins of adipocytes, a 2D- liquid chromatography/mass spectrometry/mass spectrometry protocol has been established. With the use of reversed-phase high-performance liquid chromatography, intact proteins were separated in the first dimension into eight fractions, then digested with Lys-C and trypsin. Comparative differences after drug treatment were assessed using 18O proteolytic labeling strategies. With the advent of more and more sophisticated instrumentation and data analysis tools, protocols like this one will likely become standard tools for scientists in the research fields of endocrinology, obesity, and diabetes. These protocols enable researchers to study the dynamic drug-induced changes in a comprehensive and systematic manner that was inconceivable just a few years ago.
|Original language||English (US)|
|Number of pages||10|
|Journal||Methods in molecular biology (Clifton, N.J.)|
|State||Published - Jul 31 2008|