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Improved Method for Proteome Mapping of the Liver by 2-DE MALDI-TOF MS

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journal contribution
posted on 02.04.2020, 14:57 by Giuseppe Gazzana, Jürgen Borlak
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. Keywords: Rat liver • Proteome map • 2-D PAGE • Pre-fractionation • MALDI-TOF/TOF