A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes
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We have undertaken a systematic proteomic approach to purify and identify secreted factors that are differentially expressed in preadipocytes versus adipocytes. Using one-dimensional gel electrophoresis combined with nanoelectrospray tandem mass spectrometry, proteins that were specifically secreted by 3T3-L1 preadipocytes or adipocytes were identified. In addition to a number of previously reported molecules that are up- or down-regulated during this differentiation process (adipsin, adipocyte complement-related protein 30 kDa, complement C3, and fibronectin), we identified four secreted molecules that have not been shown previously to be expressed differentially during the process of adipogenesis. Pigment epithelium-derived factor, a soluble molecule with potent antiangiogenic properties, was found to be highly secreted by preadipocytes but not adipocytes. Conversely, we found hippocampal cholinergic neurostimulating peptide, neutrophil gelatinase-associated lipocalin, and haptoglobin to be expressed highly by mature adipocytes. We also used liquid chromatography-based separation followed by automated tandem mass spectrometry to identify proteins secreted by mature adipocytes. Several additional secreted proteins including resistin, secreted acidic cysteine-rich glycoprotein/osteonectin, stromal cell-derived factor-1, cystatin C, gelsolin, and matrix metalloprotease-2 were identified by this method. To our knowledge, this is the first study to identify several novel secreted proteins by adipocytes by a proteomic approach using mass spectrometry.
|Journal||Molecular and Cellular Proteomics|
|Number of pages||9|
|Publication status||Published - 2002|
Keywords: 3T3 Cells; Acute-Phase Proteins; Adipocytes; Animals; Cell Differentiation; Eye Proteins; Gene Expression; Haptoglobins; Lipocalins; Mice; Nerve Growth Factors; Neuropeptides; Oncogene Proteins; Proteins; Proteome; Serpins; Spectrometry, Mass, Electrospray Ionization