Identification of the site of acetyl-S-enzyme formation on avian liver mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase.
Identification of the site of acetyl-S-enzyme formation on avian liver mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase.
复制标题
禽肝线粒体 3-羟基-3-甲基戊二酰辅酶 A 合酶上乙酰基-S-酶形成位点的鉴定。
DOI：
10.1021/bi00412a014
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Miziorko,HM
中科院分区：
文献类型：
--
作者：
Vollmer,SH;Mende-Mueller,LM;Miziorko,HM
Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin 53226 Received December 28, 1987; Revised Manuscript Received February 17, 1988 abstract: Avian liver mitochondrial hydroxymethylglutaryl-CoA synthase contains an active-site cysteine involved in forming the labile acetyl-S-enzyme intermediate. Identification of and assignment of function to this cysteine have been accomplished by use of an experimental strategy that relies upon generation and rapid purification of the 5-acetylcysteine-containing active-site peptide under mildly acidic conditions that stabilize the thioester adduct. Automated Edman degradation techniques indicate the peptide’s sequence to be Arg-Glu-Ser-Gly-Asn-Thr-Asp-Val-Glu-Gly-Ile-Asp-Thr-Thr-Asn-Ala-Cys-Tyr. Theacetylated cysteine corresponds to position 129 in the sequence deduced from cDNA data for the hamster cytosolic enzyme [Gil, G., Goldstein, J. L., Slaughter, C. A., & Brown, M. S.(1986) J. Biol. Chem. 261, 3710-3716], The acetyl-peptide sequence overlaps that reported for a tryptic peptide that contains a cysteine targeted by the affinity label 3-chloropropionyl-CoA [Miziorko,. M., & Behnke, C. E.(1985) J. Biol. Chem. 260, 13513-13516]. Thus, availability of these structural data allows unambiguousassignment of the acetylation site on the protein as well as a refinement of the mechanism explaining the previously observed affinity labeling of the enzyme.
