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编码大鼠磷脂酰肌醇转移蛋白的cDNA克隆的分离与测序

Isolation and sequence of cDNA clones encoding rat phosphatidylinositol transfer protein.

作者信息

Dickeson S K, Lim C N, Schuyler G T, Dalton T P, Helmkamp G M, Yarbrough L R

机构信息

Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66103.

出版信息

J Biol Chem. 1989 Oct 5;264(28):16557-64.

PMID:2777797
Abstract

Phosphatidylinositol (PtdIns) transfer protein is a cytosolic protein that catalyzes the transfer of PtdIns between membranes. It is expressed in organisms from yeast to man, and activity has been found in all animal tissues examined. Using antibodies prepared against bovine brain PtdIns transfer protein, lambda gt11 rat brain cDNA libraries were screened and several clones isolated. DNA sequence analysis showed that the cDNAs encoded a polypeptide of 271 amino acids with a mass of 31,911 Da. Comparison of the deduced amino acid sequence with N-terminal sequence data obtained for the intact purified bovine brain protein and rat lung phospholipid transfer protein verified that the cDNAs were PtdIns transfer protein clones. The predicted protein shows no significant sequence similarity to other known (phospholipid)-binding proteins. DNA blot hybridization suggests that the rat genome may contain more than one gene encoding PtdIns transfer protein. RNA blot hybridization reveals that the PtdIns transfer protein gene is expressed at low levels in a wide variety of rat tissues; all tissues examined showed a major mRNA component of 1.9 kilobases and a minor component of 3.4 kilobases. The isolation of clones encoding rat PtdIns transfer protein will greatly facilitate studies of the structure and function of PtdIns transfer proteins and their role in lipid metabolism.

摘要

磷脂酰肌醇(PtdIns)转移蛋白是一种胞质蛋白,可催化PtdIns在膜之间的转移。它在从酵母到人类的生物体中均有表达,并且在所检测的所有动物组织中都发现了其活性。利用针对牛脑PtdIns转移蛋白制备的抗体,对λgt11大鼠脑cDNA文库进行筛选并分离出了几个克隆。DNA序列分析表明,这些cDNA编码一个由271个氨基酸组成的多肽,分子量为31,911 Da。将推导的氨基酸序列与完整纯化的牛脑蛋白和大鼠肺磷脂转移蛋白的N端序列数据进行比较,证实这些cDNA是PtdIns转移蛋白克隆。预测的蛋白与其他已知的(磷脂)结合蛋白没有明显的序列相似性。DNA印迹杂交表明,大鼠基因组可能包含不止一个编码PtdIns转移蛋白的基因。RNA印迹杂交显示,PtdIns转移蛋白基因在大鼠的多种组织中低水平表达;所有检测的组织都显示出一个主要的1.9千碱基mRNA组分和一个次要的3.4千碱基组分。编码大鼠PtdIns转移蛋白的克隆的分离将极大地促进对PtdIns转移蛋白的结构和功能及其在脂质代谢中的作用的研究。

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