Evaluation of protein phosphorylation during adipogenesis

Xi Li, Rong Zeng, Qi Qun Tang

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Adipocyte differentiation is a complex process that involves the sequential expression of various adipocyte-specific genes controlled by signaling pathways and transcription factors for which phosphorylation plays a crucial regulatory role. CCAAT/enhancer-binding proteins and peroxisome proliferator-activated receptors are the most important transcriptional regulators in adipogenesis, and the functions of these proteins are regulated by various phosphorylation events. Because cultured 3T3-L1 preadipocytes are commonly used as a model for adipocyte differentiation, we used these cells for a proteomic analysis to identify kinases, phosphatases, and phosphosites that participate in adipogenesis. In addition to the phosphoproteomic analysis, we provide a detailed description of Western blotting, an in vitro phosphorylation assay, enzyme-linked immunosorbent assay, and phosphorylation site mutagenesis to fully characterize the phosphorylation of proteins and verify their roles in adipogenesis.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
Pages279-295
Number of pages17
Volume538
DOIs
StatePublished - 2014
Externally publishedYes

Publication series

NameMethods in Enzymology
Volume538
ISSN (Print)00766879
ISSN (Electronic)15577988

Keywords

  • Adipogenesis
  • Phosphoproteomic analysis
  • Phosphorylation
  • Phosphorylation site mutagenesis
  • Sandwich ELISA for phosphoprotein

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Evaluation of protein phosphorylation during adipogenesis'. Together they form a unique fingerprint.

Cite this