PMID: 11905958Mar 22, 2002Paper

A two-dimensional map and database of soluble nuclear proteins from HepG2 cells as reference for identification of nutrient-regulated transcription factors

European Journal of Nutrition
H BaumeisterW Meyerhof

Abstract

Eukaryotic cells of higher organisms are able to regulate gene transcription in response to changes in the supply of nutrients. In hepatocytes, extracellular glucose levels affect transcription of genes that encode enzymes engaged in glycolysis, gluconeogenesis and lipogenesis. While glucose response elements have been located within a few model gene promoters, the identity of glucose-sensing transcription factors and the mechanisms of their activation remain to be elucidated. We intended to establish a two-dimensional map of nuclear proteins as a reference for identification of nutrient-regulated transcription factors. Human hepatoma HepG2 cells were used for the preparation of nuclear extracts. 150-200 microg of the protein mixture were analyzed by 2-dimensional gel electrophoresis (2-DE) and silver-stained protein spots were identified by MALDI-TOF mass spectrometry. Nuclear extracts capable of transcriptional initiation and elongation and containing low amounts of cytoplasmic contaminations were prepared. 543 spots between 17 and 100 kDa and pI 3.7 and 8.8 have been resolved. From these, 65 spots were analyzed by MALDI-TOF mass spectrometry and 53 spots were identified as known proteins of which six represented transcriptio...Continue Reading

Citations

Aug 17, 2012·Public Health Nutrition·Patrícia BarbieriDaniela S Sartorelli
Dec 25, 2015·Journal of Neuroscience Methods·Urszula JankowskaSylwia Kedracka-Krok
Nov 3, 2010·Mass Spectrometry Reviews·Ganesh Kumar AgrawalRandeep Rakwal
Oct 1, 2004·Molecular & Cellular Proteomics : MCP·Ekaterina V LazDavid J Waxman
Feb 28, 2019·Revista paulista de pediatria : orgão oficial da Sociedade de Pediatria de São Paulo·Camila Silva FerreiraAna Elisa Madalena Rinaldi

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