CLONING AND SEQUENCE-ANALYSIS OF CDNA FOR HUMAN CATHEPSIN-D
CLONING AND SEQUENCE-ANALYSIS OF CDNA FOR HUMAN CATHEPSIN-D
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DOI：
10.1073/pnas.82.15.4910
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
CHIRGWIN, JM
中科院分区：
文献类型：
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作者：
FAUST, PL;KORNFELD, S;CHIRGWIN, JM
用人肾素外显子3基因组片段筛选人肝癌G2基因文库，获得1110个碱基对的人组织蛋白酶D基因克隆。用该组织蛋白D克隆进行Poly(A)+RNA印迹分析表明，该克隆的信息长度为.apprx。2.2千碱基。将该部分克隆用于筛选大小选择的人肾cDNA文库，从中筛选出2个含有apprx插入片段的组织蛋白酶D重组质粒。获得了2200和2150个碱基对。测定了这些克隆和λgt10克隆的核苷酸序列。从该序列预测的氨基酸序列显示，人组织蛋白酶D由412个氨基酸组成，其中前段和前段分别有20个和44个氨基酸。成熟蛋白区段与猪组织蛋白酶D有87%的氨基酸同源性，但不同之处在于增加了9个氨基酸。其中两个位于COOH端；另外7个位于先前确定的猪组织蛋白酶D的L链和H链的连接处之间。人组织蛋白酶D与其他天冬氨酸蛋白酶之间有高度的序列同源性，表明该蛋白家族的三维结构是保守的。
An 1110-base-pair cDNA clone for human cathepsin D was obtained by screening a .lambda.gt10 human hepatoma G2 cDNA library with a human renin exon 3 genomic fragment. Poly(A)+ RNA blot analysis with this cathepsin D clone demonstrated a message length of .apprx. 2.2 kilobases. The partial clone was used to screen a size-selected human kidney cDNA library, from which 2 cathepsin D recombinant plasmids with inserts of .apprx. 2200 and 2150 base pairs were obtained. The nucleotide sequences of these clones and of the .lambda.gt10 clone were determined. The amino acid sequence predicted from the cDNA sequence shows that human cathepsin D consists of 412 amino acids with 20 and 44 amino acids in a pre- and a prosegment, respectively. The mature protein region shows 87% amino acid identity with porcine cathepsin D but differs in having 9 additional amino acids. Two of these are at the COOH terminus; the other 7 are positioned between the previously determined junction for the L and H chains of porcine cathepsin D. A high degree of sequence homology was observed between human cathepsin D and other aspartyl proteases, suggesting a conservation of 3-dimensional structure in this family of proteins.