编码人铁螯合酶的cDNA的分子克隆及序列分析

Molecular cloning and sequence analysis of cDNA encoding human ferrochelatase.

作者信息

Nakahashi Y, Taketani S, Okuda M, Inoue K, Tokunaga R

机构信息

Third Department of Internal Medicine, Kansai Medical University, Osaka, Japan.

出版信息

Biochem Biophys Res Commun. 1990 Dec 14;173(2):748-55. doi: 10.1016/s0006-291x(05)80099-3.

DOI:10.1016/s0006-291x(05)80099-3

PMID:2260980

Abstract

The cDNA encoding human ferrochelatase [EC 4.99.1.1] was isolated from a human placenta cDNA library in bacteriophage lambda gt11 by screening with a radiolabeled fragment of mouse ferrochelatase cDNA. The cDNA had an open reading frame of 1269 base pairs (bp) encoding a protein of 423 amino acid residues (Mr. 47,833) with alternative putative polyadenylation signals in the 3' non-coding regions and poly (A) tails. Amino acid sequencing showed that the mature protein consists of 369 amino acid residues (Mr. 42,158) with a putative leader sequence of 54 amino acid residues. The human enzyme showed an 88% identity to mouse enzyme and 46% to yeast enzyme. Northern blot analysis showed two mRNAs of about 2500 and 1600 bp for ferrochelatase in K562 and HepG2 cells. As full-length cDNA for human ferrochelatase is now available, molecular lesions related to erythropoietic protoporphyria can be characterized.

摘要

通过用放射性标记的小鼠铁螯合酶cDNA片段进行筛选，从噬菌体λgt11中的人胎盘cDNA文库中分离出编码人铁螯合酶[EC 4.99.1.1]的cDNA。该cDNA具有1269个碱基对（bp）的开放阅读框，编码一个由423个氨基酸残基组成的蛋白质（Mr. 47,833），在3'非编码区有替代的假定聚腺苷酸化信号和聚（A）尾。氨基酸测序表明，成熟蛋白由369个氨基酸残基组成（Mr. 42,158），具有一个由54个氨基酸残基组成的假定前导序列。人酶与小鼠酶有88%的同一性，与酵母酶有46%的同一性。Northern印迹分析显示，在K562和HepG2细胞中铁螯合酶有两种约2500和1600 bp的mRNA。由于现在有了人铁螯合酶的全长cDNA，与红细胞生成性原卟啉症相关的分子病变就可以得到表征。