Funk C D, Rådmark O, Fu J Y, Matsumoto T, Jörnvall H, Shimizu T, Samuelsson B
Department of Physiological Chemistry, Karolinska Institutet, Stockholm, Sweden.
Proc Natl Acad Sci U S A. 1987 Oct;84(19):6677-81. doi: 10.1073/pnas.84.19.6677.
A cDNA clone corresponding to leukotriene A4 hydrolase was isolated from a human lung lambda gt11 expression library by immunoscreening with a polyclonal antiserum. Several additional clones from human lung and placenta cDNA lambda g11 libraries were obtained by plaque hybridization with the 32P-labeled lung cDNA clone. One of these clones has an insert of 1910 base pairs that contains the complete protein-coding region. From the deduced primary structure, leukotriene A4 hydrolase is a 610 amino and protein with a calculated molecular weight of 69,140. No apparent homologies with microsomal epoxide hydrolases were found. RNA blot analysis indicated substantial amounts of a discrete mRNA of approximately equal to 2250 nucleotides in lung tissue and leukocytes.
通过用多克隆抗血清进行免疫筛选,从人肺λgt11表达文库中分离出一个与白三烯A4水解酶对应的cDNA克隆。通过用32P标记的肺cDNA克隆进行噬菌斑杂交,从人肺和胎盘cDNAλg11文库中获得了几个额外的克隆。其中一个克隆有一个1910个碱基对的插入片段,包含完整的蛋白质编码区。从推导的一级结构来看,白三烯A4水解酶是一种由610个氨基酸组成的蛋白质,计算分子量为69140。未发现与微粒体环氧化物水解酶有明显的同源性。RNA印迹分析表明,肺组织和白细胞中存在大量约2250个核苷酸的离散mRNA。