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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 
 JOURNAL OF PROTEOME RESEARCH, Vol.8(1) : 48-58, 2009 
Journal Title
Issue Date
3T3-L1 Cells ; Adipocytes/metabolism* ; Adipogenesis ; Animals ; Azo Compounds/pharmacology ; Cell Differentiation ; Cell Nucleus/metabolism ; Chromatography, Liquid/methods ; Gene Expression Regulation* ; Isotopes ; Mass Spectrometry/methods ; Mice ; Proteomics/methods* ; Time Factors ; Trypsin/pharmacology
Adipocyte ; adipogenesis ; proteomics ; SILAC
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.
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1. College of Medicine (의과대학) > Dept. of Biochemistry and Molecular Biology (생화학-분자생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Jae Woo(김재우) ORCID logo https://orcid.org/0000-0001-5456-9495
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