从大鼠肝脏中S-腺苷-L-高半胱氨酸水解酶的cDNA序列推导出来的氨基酸序列。
Amino acid sequence of S-adenosyl-L-homocysteine hydrolase from rat liver as derived from the cDNA sequence.
作者信息
Ogawa H, Gomi T, Mueckler M M, Fujioka M, Backlund P S, Aksamit R R, Unson C G, Cantoni G L
出版信息
Proc Natl Acad Sci U S A. 1987 Feb;84(3):719-23. doi: 10.1073/pnas.84.3.719.
Abstract
Rat liver cDNA libraries constructed in lambda gt11 were screened for reactivity with polyclonal antibodies to native S-adenosyl-L-homocysteine (AdoHcy) hydrolase (adenosylhomocysteinase; EC 3.3.1.1). Five clones were isolated and sequenced. The amino acid sequence, deduced from the cDNA sequence, contained the sequence of eight peptides obtained by tryptic and cyanogen bromide fragmentation of rat liver AdoHcy hydrolase. Identification of the amino- and carboxyl-terminal peptides in the amino acid sequence showed that the complete sequence was obtained. A "fingerprint" sequence was found that is characteristic of dinucleotide-binding domains of many proteins. For AdoHcy hydrolase, this region from the lysine at position 213 to the aspartate at position 244, containing the sequence Gly-Xaa-Gly-Xaa-Xaa-Gly at positions 219-224, is presumably the site of binding for NAD+, which is required for the activity of the enzyme.
摘要
用λgt11构建的大鼠肝脏cDNA文库，针对与天然S-腺苷-L-高半胱氨酸（AdoHcy）水解酶（腺苷高半胱氨酸酶；EC 3.3.1.1）的多克隆抗体的反应性进行筛选。分离出五个克隆并进行测序。从cDNA序列推导的氨基酸序列，包含了通过大鼠肝脏AdoHcy水解酶的胰蛋白酶和溴化氰片段化获得的八个肽段的序列。氨基酸序列中氨基末端和羧基末端肽段的鉴定表明获得了完整序列。发现了一个“指纹”序列，它是许多蛋白质二核苷酸结合结构域的特征。对于AdoHcy水解酶，从第213位的赖氨酸到第244位的天冬氨酸的这个区域，在第219 - 224位包含序列Gly-Xaa-Gly-Xaa-Xaa-Gly，大概是NAD + 的结合位点，而NAD + 是该酶活性所必需的。
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