Quinn J, Diamond A G, Masters A K, Brookfield D E, Wallis N G, Yeaman S J
Department of Biochemistry and Genetics, Medical School, University of Newcastle upon Tyne, U.K.
Biochem J. 1993 Jan 1;289 ( Pt 1)(Pt 1):81-5. doi: 10.1042/bj2890081.
The dihydrolipoamide acetyltransferase subunit (E2p) of mammalian pyruvate dehydrogenase complex has two highly conserved lipoyl domains each modified with a lipoyl cofactor bound in amide linkage to a specific lysine residue. A sub-gene encoding the inner lipoyl domain of human E2p has been over-expressed in Escherichia coli. Two forms of the domain have been purified, corresponding to lipoylated and non-lipoylated species. The apo-domain can be lipoylated in vitro with partially purified E. coli lipoate protein ligase, and the lipoylated domain can be reductively acetylated by human E1p (pyruvate dehydrogenase). Availability of the two forms will now allow detailed biochemical and structural studies of the human lipoyl domains.
哺乳动物丙酮酸脱氢酶复合体的二氢硫辛酰胺乙酰转移酶亚基（E2p）有两个高度保守的硫辛酰结构域，每个结构域都通过酰胺键与一个特定赖氨酸残基结合的硫辛酰辅因子进行修饰。编码人E2p内部硫辛酰结构域的一个亚基因已在大肠杆菌中过表达。已纯化出该结构域的两种形式，分别对应硫辛酰化和非硫辛酰化的物种。脱辅基结构域可在体外被部分纯化的大肠杆菌硫辛酸蛋白连接酶硫辛酰化，硫辛酰化结构域可被人E1p（丙酮酸脱氢酶）进行还原乙酰化。这两种形式的可得性将使得对人硫辛酰结构域进行详细的生化和结构研究成为可能。
