tailieunhanh - Báo cáo Y học: Ontogeny and subcellular localization of rat liver mitochondrial branched chain amino-acid aminotransferase

Departamento de Fisiologıa de la Nutricion, Instituto Nacional de Ciencias Medicas y Nutricion ‘Salvador Zubiran’, Mexico; ´ ´ ´ ´ ´ Departamento de Patologıa Experimental, Instituto Nacional de Ciencias Medicas y Nutricion ‘Salvador Zubiran’,Mexico; 3Department of Biochemistry, Wake Forest University Medical Center, Winston-Salem, North Carolina, USA Branched chain amino-acid aminotransferase (BCAT) activity is present in fetal liver but the developmental pattern of mitochondrial BCAT (BCATm) expression in rat liver has not been studied. The aim of this study was to determine the activity, protein and mRNA concentration of BCATm in fetal and postnatal rat liver, and to localize this enzyme at. | Eur. J. Biochem. 268 6132-6139 2001 FEBS 2001 Ontogeny and subcellular localization of rat liver mitochondrial branched chain amino-acid aminotransferase Nimbe Torres1 Carolina Varaas1 Roaelio Hernandez-Pando2 Hector Orozco2 Susan M. Hutson3 and BBB. Armando R. Tovar1 1 Departamento de Fisiologia de la Nutricion Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran Mexico 2Departamento de Patologia Experimental InstitutoNacional de Ciencias Medicas y Nutricion Salvador Zubiran Mexico 3Department of Biochemistry Wake Forest University Medical Center Winston-Salem North Carolina USA Branched chain amino-acid aminotransferase BCAT activity is present in fetal liver but the developmental pattern of mitochondrial BCAT BCATm expression in rat liver has not been studied. The aim of this study was to determine the activity protein and mRNA concentration of BCATm in fetal and postnatal rat liver and to localize this enzyme at the cellular and subcellular levels at both developmental stages. Maximal BCAT activity and BCATm mRNA expression occurred at 17 days gestation in fetal rat liver and then declined significantly immediately after birth. This pattern was observed only in liver rat heart showed a different developmental pattern. Fetal liver showed intense immunostaining to BCATm in the nuclei and mitochondria of hepatic cells and blood cell precursors in contrast adult liver showed mild immunoreactivity located only in the mitochondria of hepatocytes. BCAT activity in isolated fetal liver nuclei was mU-mg21 protein whereas it was undetectable in adult liver nuclei. By Western blot analysis the BCATm antibody recognized a 41-kDa protein in fetal liver nuclei and proteins of 41 and 43 kDa in fetal liver supernatant. In adult rat liver supernatant the BCATm antibody recognized only a 43-kDa protein however neither protein was detected in adult rat liver nuclei. The appearance of the 41-kDa protein was associated with the presence of the highly active form

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