Improved method for proteome mapping of the liver by 2-DE MALDI-TOF MS.
Identifying the liver proteome has been the subject of intensified research. Notably, 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 identifiable proteins. In our efforts, we used two different lysis buffers in sequence, a liquid-phase IEF pre-fractionation and separation of protein mixtures at three different pH ranges (3-10, 5-8, and 7-10). Then, >15 000 gel digested proteins were investigated. We report an identification of 590 different gene products, including some isoforms. More than 150 proteins have not been reported so far by two-dimensional electrophoresis (2-DE) proteome mapping. We further studied the transcript expression of more than 33 000 genes in rat liver to explore correlations between transcript and protein expression. Overall, we report a method for the separation of rat liver proteins and their identification by mass spectrometry. The newly identified proteins will enable an improved understanding of liver biology.
Gazzana
Giuseppe
G
Department of Molecular Medicine and Medical Biotechnology, Fraunhofer Institute of Toxicology and Experimental Medicine, Nikolai-Fuchs-Strasse 1, 30625 Hannover, Germany.
Borlak
Jürgen
J
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008110:Liver Extracts / Q000032:analysis*; D008297:Male; D020543:Proteome / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D008110:Liver Extracts; D020543:Proteome