Heterologously expressed inner lipoyl domain of dihydrolipoyl acetyltransferase inhibits ATP-dependent inactivation of pyruvate dehydrogenase complex - Identification of important amino acid residues

被引:24
作者
Jackson, JC
Vinluan, CC
Dragland, CJ
Sundararajan, V
Yan, B
Gounarides, JS
Nirmala, NR
Topiol, S
Ramage, P
Blume, JE
Aicher, TD
Bell, PA
Mann, WR
机构
[1] Novartis Inst Biomed Res, Metab & Cardiovasc Dis Res, Summit, NJ 07901 USA
[2] Novartis Pharma Ltd, Biomol Prod Unit, CH-4002 Basel, Switzerland
关键词
D O I
10.1042/bj3340703
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The activity of the pyruvate dehydrogenase multienzyme complex (PDC), which catalyses the oxidation of pyruvate to acetyl-CoA within the mitochondrion, is diminished in animal models of diabetes, Studies with purified PDC components have suggested that the kinases responsible for inactivating the decarboxylase catalytic subunits of the complex are most efficient in their regulatory role when they are bound to dihydrolipoyl acetyltransferase (E2) subunits, which form the structural core of the complex. We report that the addition of an exogenous E2 subdomain (inner lipoyl domain) to an intact PDC inhibits ATP-dependent inactivation of the complex. By combining molecular modelling, site-directed mutagenesis and biophysical characterizations, we have also identified two amino acid residues in this subdomain (IIe(229) and Phe(231)) that largely determine the magnitude of this effect.
引用
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页码:703 / 711
页数:9
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