Naggert J, Witkowski A, Mikkelsen J, Smith S
Children's Hospital Oakland Research Institute, California 94609.
J Biol Chem. 1988 Jan 25;263(3):1146-50.
A cloned cDNA containing the entire coding sequence for the long-chain S-acyl fatty acid synthetase thioester hydrolase (thioesterase I) component as well as the 3'-noncoding region of the fatty acid synthetase has been isolated using an expression vector and domain-specific antibodies. The coding region was assigned to the thioesterase I domain by identification of sequences coding for characterized peptide fragments, amino-terminal analysis of the isolated thioesterase I domain and the presence of the serine esterase active-site sequence motif. The thioesterase I domain is 306 amino acids long with a calculated molecular mass of 33,476 daltons; its DNA is flanked at the 5'-end by a region coding for the acyl carrier protein domain and at the 3'-end by a 1,537-base pairs-long noncoding sequence with a poly(A) tail. The thioesterase I domain exhibits a low, albeit discernible, homology with the discrete medium-chain S-acyl fatty acid synthetase thioester hydrolases (thioesterase II) from rat mammary gland and duck uropygial gland, suggesting a distant but common evolutionary ancestry for these proteins.
利用表达载体和结构域特异性抗体,已分离出一个克隆的互补DNA(cDNA),其包含长链S-酰基脂肪酸合成酶硫酯水解酶(硫酯酶I)组分的完整编码序列以及脂肪酸合成酶的3'-非编码区。通过鉴定编码特定肽片段的序列、对分离出的硫酯酶I结构域进行氨基末端分析以及存在丝氨酸酯酶活性位点序列基序,将编码区定位到硫酯酶I结构域。硫酯酶I结构域由306个氨基酸组成,计算分子量为33476道尔顿;其DNA在5'-端侧翼是编码酰基载体蛋白结构域的区域,在3'-端侧翼是一个1537个碱基对长的带有聚腺苷酸尾巴的非编码序列。硫酯酶I结构域与来自大鼠乳腺和鸭尾脂腺的离散中链S-酰基脂肪酸合成酶硫酯水解酶(硫酯酶II)表现出低但可辨别的同源性,这表明这些蛋白质有着遥远但共同的进化起源。