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A map of the rat liver proteome

Abstract 35.4
: Gazzana, G.; Borlak, J.

Molecular & cellular proteomics 4 (2005), Nr.8, Supplement, S.S339
ISSN: 1535-9476
Fraunhofer ITEM ()
Rats; Liver; Proteome

The number of proteins expressed in eukaryotic cells may vary between several thousands to half a million. Single hepatic cells express on average about 10,000 different proteins. Due to their strong heterogeneity in size, charge, solubility and relative abundance, different strategies of pre-fractionation must be employed to increase the number of detectable proteins. For proteome mapping, liver extracts of Rattus Norvegicus were separated by two-dimensional SDS-PAGE. We generated to the best of our knowledge the largest proteome 2-DE PAGE map of rat liver so far (review in progress), by employing two different lysis buffers in sequence, a pre-IEF, and three different pH ranges (3-10, 5-8 and 7-10). In the past, Fountoulakis et al., (Journal of Chromatography B, 782 (2002) 197-218) reported 273 hepatic proteins, while Kromer et al., identified 114 proteins of the rat liver proteome (Proteomics, 3, 2003, 1835-1862). We now report 560 different gene products, including several isoforms. More than 150 proteins are novel and include the contraception-associated protein 1 (CAP1), a low abundance ubiquitous protein which has been classified as a oncogene (Wagenfeld, A. et al., J Androl. 2000 Nov-Dex; 21(6):954-63), as well as hepatorma subtracted clone one (HSCO), a mitochondrial protein that accelerates export of NF-kappa B from the nucleus and inhibits p53-dependent apoptosis (Higashitsuji et al., Cancer Cell. 2002 Oct; 2(4):335-46). We investigated the correlation between gene expression, as determined by oligo microchip analysis of more than 33,000 gene, and the proteins identified from 2-DE PAGE/MALDI-TOF-TOF. The overall correlation was very good indeed, e.g. 83 %.