caa a22 4500 397573278 CHVBK 20180308164847.0 cr unu---uuuuu 161202e199610 xx s 000 0 eng 10.1093/hmg/5.10.1643 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.10.1643 Functional Analysis in Saccharomyces Cerevisiae of Naturally Occurring Amino Acid Substitutions in Human Dihydrolipoamide Dehydrogenase [Elektronische Daten] [Margaret M. Lanterman, J. Richard Dickinson, Dean J. Danner] Dihydrolipoamide dehydrogenase is a common component of mammalian multienzyme complexes that decarboxylate α-ketoacids and catabolize glycine. The common function is to reoxidize a reduced lipoate component of each complex, thereby preparing that lipoate for another round of catalysis. Regions within dihydrolipoamide dehydrogenase involved in association with other proteins of the complexes are poorly defined, and despite high amino acid sequence conservation through evolution, it is unknown if dihydro-lipoamide dehydrogenases are functionally equivalent across species. To address this issue, we asked whether the human enzyme could restore function to the -ketoacid dehydrogenase complexes in a yeast strain deficient in endogenous dihydrolipoamide dehydro-genase. This dihydrolipoamide dehydrogenase null mutant will not grow on non-fermentable carbon sources. The human enzyme expressed from a CEN plasmid complemented the growth phenotype and restored full activity to the pyruvate and α-ketoglutarate dehydrogenase complexes. Human dihydrolipoamide dehydrogenases with selected amino acid substitutions were then tested in the null strain for their ability to restore function. Substitutions tested represented naturally occurring candidate mutations identified in an individual with inactive dihydrolipoamide dehydro-genase. A K37E change had full function while a P453L change resulted in reduced dihydrolipoamide dehydrogenase abundance in the mitochondria and no detectable catalytic activity. © 1996 Oxford University Press Lanterman Margaret M. Department of Genetics and Molecular Medicine, Emory University School of Medicine, 1462 Clifton Road, Atlanta, GA 30322, USA aut Dickinson J. Richard School of Pure and Applied Biology, University of Wales, College of Cardiff, Cardiff CF1 3TL, UK aut Danner Dean J. Department of Genetics and Molecular Medicine, Emory University School of Medicine, 1462 Clifton Road, Atlanta, GA 30322, USA aut Human Molecular Genetics Oxford University Press 5/10(1996-10), 1643-1648 0964-6906 5:10<1643 1996 5 hmg https://doi.org/10.1093/hmg/5.10.1643 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.10.1643 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lanterman Margaret M. Department of Genetics and Molecular Medicine, Emory University School of Medicine, 1462 Clifton Road, Atlanta, GA 30322, USA aut NATIONALLICENCE 700 1- Dickinson J. Richard School of Pure and Applied Biology, University of Wales, College of Cardiff, Cardiff CF1 3TL, UK aut NATIONALLICENCE 700 1- Danner Dean J. Department of Genetics and Molecular Medicine, Emory University School of Medicine, 1462 Clifton Road, Atlanta, GA 30322, USA aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/10(1996-10), 1643-1648 0964-6906 5:10<1643 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford