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小鼠四聚体羰基还原酶的克隆、表达及组织分布。与一种脂肪细胞27 kDa蛋白的同一性。

Cloning, expression and tissue distribution of mouse tetrameric carbonyl reductase. Identity with an adipocyte 27-kDa protein.

作者信息

Nakanishi M, Deyashiki Y, Ohshima K, Hara A

机构信息

Laboratory of Biochemistry, Gifu Pharmaceutical University, Mitahora-higashi, Japan.

出版信息

Eur J Biochem. 1995 Mar 1;228(2):381-7.

PMID:7705352
Abstract

We previously cloned a cDNA for pig lung tetrameric carbonyl reductase which shows significant similarity to a putative 27-kDa protein predicted from the cDNA for murine adipocyte RNA which had been increased in its differentiation [Nakanishi, M., Deyashiki, Y., Nakayama, T., Sato, K. & Hara, A. (1993) Biochem. Biophys. Res. Commun. 194, 1311-1316]. In this investigation, we isolated and sequenced a full-length cDNA for the tetrameric enzyme from a mouse lung cDNA library. It consisted of 984 bp and coded for a protein of 244 amino acid residues, of which 202 residues were identified by protein sequencing. The expression of the cDNA in Escherichia coli resulted in synthesis of a protein structurally and functionally similar to the enzyme purified from mouse lung. The nucleotide sequence of the cDNA was virtually identical to that of the cDNA for the adipocyte 27-kDa protein. Although Northern-blot analysis of mouse tissues showed that enzyme mRNA to be 1.1 kb only in lung, low expression of the mRNA in all the extrapulmonary tissues, including adipose tissue, was demonstrated by a reverse-transcription PCR method. Western-blot analysis also indicated the presence of the enzyme in the adipose tissue. This is the first report on an identification of the putative gene product of adipocytes as tetrameric carbonyl reductase, the expression of which is tissue-specifically regulated.

摘要

我们之前克隆了猪肺四聚体羰基还原酶的cDNA,它与从小鼠脂肪细胞RNA的cDNA预测的一种假定的27 kDa蛋白具有显著相似性,该蛋白在其分化过程中有所增加[Nakanishi, M., Deyashiki, Y., Nakayama, T., Sato, K. & Hara, A. (1993) Biochem. Biophys. Res. Commun. 194, 1311 - 1316]。在本研究中,我们从小鼠肺cDNA文库中分离并测序了该四聚体酶的全长cDNA。它由984 bp组成,编码一个含有244个氨基酸残基的蛋白质,其中202个残基通过蛋白质测序得以鉴定。该cDNA在大肠杆菌中的表达导致合成了一种在结构和功能上与从小鼠肺中纯化的酶相似的蛋白质。该cDNA的核苷酸序列与脂肪细胞27 kDa蛋白的cDNA序列几乎相同。虽然对小鼠组织的Northern印迹分析表明该酶的mRNA仅在肺中为1.1 kb,但逆转录PCR方法证明该mRNA在包括脂肪组织在内的所有肺外组织中表达水平较低。蛋白质印迹分析也表明脂肪组织中存在该酶。这是关于将脂肪细胞假定的基因产物鉴定为四聚体羰基还原酶的首次报道,其表达受到组织特异性调控。

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