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人类嗜酸性粒细胞过氧化物酶染色体基因的分子克隆与特性分析

Molecular cloning and characterization of a chromosomal gene for human eosinophil peroxidase.

作者信息

Sakamaki K, Tomonaga M, Tsukui K, Nagata S

机构信息

Osaka Bioscience Institute, Japan.

出版信息

J Biol Chem. 1989 Oct 5;264(28):16828-36.

PMID:2550461
Abstract

Using human myeloperoxidase cDNA as a probe, a chromosomal gene related to myeloperoxidase was isolated from a human gene library. Comparison of the amino acid sequence deduced from the nucleotide sequence of the cloned gene with that of human eosinophil peroxidase purified from buffy coats has indicated that the isolated gene is the chromosomal gene for human eosinophil peroxidase. Like human myeloperoxidase gene, human eosinophil peroxidase gene consists of 12 exons and 11 introns spanning about 12 kilobases. The gene can code for a protein of 715 amino acids with a calculated Mr of 81,036. The heavy chain and the light chain of eosinophil peroxidase were located on the COOH and NH2 terminus of the protein, respectively. The coding sequences of eosinophil peroxidase and myeloperoxidase show homologies of 72.4% at the nucleotide and 69.8% at the amino acid level, while little homology was found in the 5'-flanking region. Northern hybridization and S1 mapping analysis of RNA from human leukemic cells have indicated that the eosinophil peroxidase gene is expressed in the eosinophilic subline of human HL-60 cells but not in the neutrophilic subline or in parental HL-60 cells.

摘要

以人髓过氧化物酶cDNA为探针,从人基因文库中分离出一个与髓过氧化物酶相关的染色体基因。将从克隆基因的核苷酸序列推导的氨基酸序列与从血沉棕黄层中纯化的人嗜酸性粒细胞过氧化物酶的氨基酸序列进行比较,结果表明所分离的基因是人类嗜酸性粒细胞过氧化物酶的染色体基因。与人髓过氧化物酶基因一样,人嗜酸性粒细胞过氧化物酶基因由12个外显子和11个内含子组成,跨越约12千碱基。该基因可编码一种由715个氨基酸组成的蛋白质,计算的分子量为81,036。嗜酸性粒细胞过氧化物酶的重链和轻链分别位于该蛋白质的COOH端和NH2端。嗜酸性粒细胞过氧化物酶和髓过氧化物酶的编码序列在核苷酸水平上的同源性为72.4%,在氨基酸水平上为69.8%,而在5'侧翼区几乎没有同源性。对人白血病细胞RNA的Northern杂交和S1图谱分析表明,嗜酸性粒细胞过氧化物酶基因在人HL-60细胞的嗜酸性亚系中表达,但在嗜中性亚系或亲代HL-60细胞中不表达。

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