编码鸡卵黄氨肽酶Ey的cDNA的分离与鉴定

Isolation and characterization of cDNA encoding chicken egg yolk aminopeptidase Ey.

作者信息

Midorikawa T, Abe R, Yamagata Y, Nakajima T, Ichishima E

机构信息

Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, Miyagi, Japan.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 1998 Mar;119(3):513-20. doi: 10.1016/s0305-0491(98)00012-1.

DOI:10.1016/s0305-0491(98)00012-1

PMID:9734335

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7172579/

Abstract

Aminopeptidase Ey (EC 3.4.11.20) from chicken (Gallus gallus domesticus) egg yolk is a homodimeric exopeptidase with a broad specificity for N-terminal amino acid residues at P1 position of the substrate. Aminopeptidase Ey is a 300-k metalloexopeptidase, containing 1.0 g atom of zinc per mole of a subunit with a relative molecular mass of 150 k. A full-length cDNA was cloned from chicken (female) liver cDNA library. Analysis of the 3196-base pairs (bp) nucleotide sequence of the cDNA revealed a single open reading frame coding for 967 amino acid residues. The coding region of aminopeptidase Ey gene, apdE, occupies 2901 bp of the cDNA. The predicted amino acid sequence of the enzyme is 66, 65, 64 and 63% identical with those of aminopeptidases N (EC 3.4.11.2) from human, pig, rabbit and rat, respectively. Aminopeptidase Ey contains the metallo-binding sequence motif, His-Glu-Xaa-His, found in zinc metallopeptidases. Zinc binding sites, His-386, His-390 and Glu-409, and catalytic site, Glu-387, were conserved in the homologous aminopeptidases N.

摘要

来自家鸡（Gallus gallus domesticus）蛋黄的氨肽酶Ey（EC 3.4.11.20）是一种同二聚体外肽酶，对底物P1位置的N端氨基酸残基具有广泛的特异性。氨肽酶Ey是一种300-k的金属外肽酶，每摩尔相对分子质量为150 k的亚基含有1.0克原子的锌。从鸡（雌性）肝脏cDNA文库中克隆出全长cDNA。对该cDNA的3196个碱基对（bp）核苷酸序列分析显示有一个单一的开放阅读框，编码967个氨基酸残基。氨肽酶Ey基因apdE的编码区占cDNA的2901 bp。该酶预测的氨基酸序列与来自人、猪、兔和大鼠的氨肽酶N（EC 3.4.11.2）的氨基酸序列分别有66%、65%、64%和63%的同一性。氨肽酶Ey含有锌金属肽酶中发现的金属结合序列基序His-Glu-Xaa-His。锌结合位点His-386、His-390和Glu-409以及催化位点Glu-387在同源氨肽酶N中保守。