Springer Protocols: Abstract: 10. Applications of Proteomics to the Study of Adipose Tissue

10. Applications of Proteomics to the Study of Adipose Tissue

By: Sonja Hess³, Xiaoli Chen⁴

Abstract

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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.

Affiliation(s): (3) Proteomics and Mass Spectrometry Facility, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland
(4) Department of Food Science and Nutrition, University of Minnesota, St. Paul, Minnesota

Book Title: Adipose Tissue Protocols

Series: Methods in Molecular Biology | Volume: 456 | Pub. Date: Jun-01-2008 | Page Range: 131-140 | DOI: 10.1007/978-1-59745-245-8_10

Subject: Molecular Medicine

Key Words: Adipose tissue - quantitative proteomics - 18O-labeling - diabetes - mass spectrometry - electrospray

References

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