Temporal profiling of the adipocyte proteome during differentiation using a five-plex SILAC based strategy

Henrik Molina, Yi Yang, Travis Ruch, Jae Woo Kim, Peter Mortensen, Tamara Otto, Anuradha Nalli, Qi Qun Tang, M. Daniel Lane, Raghothama Chaerkady, Akhilesh Pandey

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

The adipose tissue has important secretory and endocrine functions in humans. The regulation of adipocyte differentiation has been actively pursued using transcriptomic methods over the last several years. Quantitative proteomics has emerged as a promising approach to obtain temporal profiles of biological processes such as differentiation. Stable isotope labeling with amino acids in cell culture (SILAC) is a simple and robust method for labeling proteins in vivo. Here, we describe the development and application of a five-plex SILAC experiment using four different heavy stable isotopic forms of arginine to study the nuclear proteome and the secretome during the course of adipocyte differentiation. Tandem mass spectrometry analysis using a quadrupole time-of-flight instrument resulted in identification of a total 882 proteins from these two proteomes. Of these proteins, 427 were identified on the basis of one or more arginine-containing peptides that allowed quantitation. In addition to previously reported molecules that are differentially expressed during the process of adipogenesis (e.g., adiponectin and lipoprotein lipase), we identified several proteins whose differential expression during adipocyte differentiation has not been documented previously. For example, THO complex 4, a context-dependent transcriptional activator in the T-cell receptor alpha enhancer complex, showed highest expression at middle stage of adipogenesis, while SNF2 alpha, a chromatin remodeling protein, was downregulated upon initiation of adipogenesis and remained so during subsequent time points. This study using a 5-plex SILAC to investigate dynamics illustrates the power of this approach to identify differentially expressed proteins in a temporal fashion.

Original languageEnglish (US)
Pages (from-to)48-58
Number of pages11
JournalJournal of Proteome Research
Volume8
Issue number1
DOIs
StatePublished - Jan 1 2009
Externally publishedYes

Fingerprint

Proteome
Adipocytes
Adipogenesis
Proteins
Labeling
Arginine
Isotope Labeling
Biological Phenomena
Chromatin Assembly and Disassembly
Lipoprotein Lipase
Adiponectin
T-Cell Antigen Receptor
Tandem Mass Spectrometry
Cell culture
Isotopes
Proteomics
Chromatin
Mass spectrometry
Adipose Tissue
Down-Regulation

Keywords

  • Adipocyte
  • Adipogenesis
  • Proteomics
  • SILAC

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Temporal profiling of the adipocyte proteome during differentiation using a five-plex SILAC based strategy. / Molina, Henrik; Yang, Yi; Ruch, Travis; Kim, Jae Woo; Mortensen, Peter; Otto, Tamara; Nalli, Anuradha; Tang, Qi Qun; Daniel Lane, M.; Chaerkady, Raghothama; Pandey, Akhilesh.

In: Journal of Proteome Research, Vol. 8, No. 1, 01.01.2009, p. 48-58.

Research output: Contribution to journalArticle

Molina, H, Yang, Y, Ruch, T, Kim, JW, Mortensen, P, Otto, T, Nalli, A, Tang, QQ, Daniel Lane, M, Chaerkady, R & Pandey, A 2009, 'Temporal profiling of the adipocyte proteome during differentiation using a five-plex SILAC based strategy', Journal of Proteome Research, vol. 8, no. 1, pp. 48-58. https://doi.org/10.1021/pr800650r
Molina, Henrik ; Yang, Yi ; Ruch, Travis ; Kim, Jae Woo ; Mortensen, Peter ; Otto, Tamara ; Nalli, Anuradha ; Tang, Qi Qun ; Daniel Lane, M. ; Chaerkady, Raghothama ; Pandey, Akhilesh. / Temporal profiling of the adipocyte proteome during differentiation using a five-plex SILAC based strategy. In: Journal of Proteome Research. 2009 ; Vol. 8, No. 1. pp. 48-58.
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