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Metabolomics

Dietary Protein–Related Changes in Hepatic Transcription Correspond to Modifications in Hepatic Protein Expression in Growing Pigs¹

Peter Junghans, Thilo Kaehne, Manfred Beyer, Cornelia C. Metges and Manfred Schwerin^*,2

Research Units Nutritional Physiology "Oskar Kellner" and ^* Molecular Biology, Research Institute for the Biology of Farm Animals Dummerstorf, D-18196 Dummerstorf, Germany and Institute of Experimental Internal Medicine, Research Center Immunology, Otto-von-Guericke-University Magdeburg, D-39120 Magdeburg, Germany

²To whom correspondence should be addressed. E-mail: schwerin{at}fbn-dummerstorf.de.

In a previous investigation we showed by expression profiling based on transcription analysis using differential display RT-PCR (DDRT-PCR) and real-time RT-PCR that a soy protein diet (SPI) significantly changes the hepatic transcription pattern compared with a casein diet (CAS). The present study was conducted to determine whether the transcriptional modulation is translated into protein expression. The hepatic mRNA abundance of four genes (EP24.16, LC3, NPAP60L, RFC2) that showed diet-related expression in previous DDRT-PCR experiments was analyzed by real-time RT-PCR. Two pigs that showed the most prominent SPI-related changes of transcription and two casein-fed pigs were selected and their hepatic protein pattern was studied comparatively by two-dimensional gel electrophoresis and peptide mass fingerprinting. The two-dimensional protein gel electrophoresis revealed a predominant SPI-associated upregulation of protein expression that corresponded to the results of the mRNA study. Of 380 diet-related protein spots displayed, 215 appeared exclusively or enlarged in the two SPI pigs; 10 of 39 diet-related expressed protein spots extracted could be identified by peptide mass fingerprinting and database search. Compared with the transcriptomics approach, the proteomics approach led in part to the identification of the same diet-associated expressed molecules (plasminogen, trypsin, phospholipase A₂, glutathione-S-transferase , retinal binding protein) or at least molecules belonging to the same metabolic pathways (protein and amino acid metabolism, oxidative stress response, lipid metabolism). The present results at the proteome level confirm SPI-related increased oxidative stress response and significant effects on protein biosynthesis already observed at the transcriptome level.

KEY WORDS: • protein diets • transcriptome • proteome • liver • swine

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