Characterization of the human adipocyte proteome and reproducibility of protein abundance by one-dimensional gel electrophoresis and HPLC - ESI - MS/MS

Xitao Xie, Zhengping Yi, Benjamin Bowen, Cassandra Wolf, Charles R. Flynn, Sandeep Sinha, Lawrence J. Mandarino, Christian Meyer

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

Abnormalities in adipocytes play an important role in various conditions, including the metabolic syndrome, type 2 diabetes mellitus and cardiovascular disease, but little is known about alterations at the protein level. We therefore sought to (1) comprehensively characterize the human adipocyte proteome for the first time and (2) demonstrate feasibility of measuring adipocyte protein abundances by one-dimensional SDS-PAGE and high performance liquid chromatography-electron spray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). In adipocytes isolated from ∼0.5 g of subcutaneous abdominal adipose tissue of three healthy, lean subjects, we identified a total of 1493 proteins. Triplicate analysis indicated a 22.5% coefficient of variation of protein abundances. Proteins ranged from 5.8 to 629 kDa and included a large number of proteins involved in lipid metabolism, such as fatty acid transport, fatty acid oxidation, lipid storage, lipolysis, and lipid droplet maintenance. Furthermore, we found most glycolysis enzymes and numerous proteins associated with oxidative stress, protein synthesis and degradation as well as some adipokines. 22% of all proteins were of mitochondrial origin. These results provide the first detailed characterization of the human adipocyte proteome, suggest an important role of adipocyte mitochondria, and demonstrate feasibility of this approach to examine alterations of adipocyte protein abundances in human diseases.

Original languageEnglish (US)
Pages (from-to)4521-4534
Number of pages14
JournalJournal of Proteome Research
Volume9
Issue number9
DOIs
StatePublished - Sep 3 2010
Externally publishedYes

Keywords

  • adipocyte
  • human
  • mitochondria
  • proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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