CLONING AND CHARACTERIZATION OF THE MITOCHONDRIAL PHOSPHATE-TRANSPORT PROTEIN GENE FROM THE YEAST SACCHAROMYCES-CEREVISIAE
CLONING AND CHARACTERIZATION OF THE MITOCHONDRIAL PHOSPHATE-TRANSPORT PROTEIN GENE FROM THE YEAST SACCHAROMYCES-CEREVISIAE
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DOI：
10.1021/bi00215a035
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发表时间：
1991-01-08
期刊：
影响因子：
2.9
通讯作者：
WOHLRAB, H
中科院分区：
文献类型：
--
作者：
PHELPS, A;SCHOBERT, CT;WOHLRAB, H
我们已经克隆了酿酒酵母磷酸盐转运蛋白（PTP）的基因，线粒体阴离子转运蛋白基因家族的成员。 由于PTP具有封闭的N-末端，我们制备了三个肽。 寡核苷酸，基于它们的序列，用于筛选Yep 24容纳的基因组文库。 克隆Y22的总共2073个碱基编码311个氨基酸的蛋白质（M（r）32814），其与阴离子转运蛋白具有相似性：一个三重基因结构和6个疏水片段。 典型的PTP，三重基因结构具有X-Pro-X-（Asp/Glu）-X-X-（Lys/Arg）-X-（Arg/Lys）-X（X是未指明的氨基酸）基序和仅在第一和第二重复之间的非常高的同源性。 这6个疏水片段含有酵母蛋白和牛肉蛋白之间保守的116个氨基酸中的大部分。 在牛肉蛋白中发现的N-末端延伸的信号序列不存在。 酵母蛋白比牛肉蛋白少33%的碱性和酸性氨基酸，少5个半胱氨酸残基。 该蛋白质对N-乙基马来酰亚胺不敏感，因为Cys-42（牛肉）已被替换为Thr。 汞磺酰敏感性一直保持，必须是由于它的三个半胱氨酸之一。 在这三个半胱氨酸中，只有位于第一疏水区段的Cys-28在酵母和牛肉蛋白之间是保守的。
We have cloned the gene of the Saccharomyces cerevisiae phosphate transport protein (PTP), a member of the mitochondrial anion transport protein gene family. As PTP has a blocked N-terminus, we prepared three peptides. Oligonucleotides, based on their sequences, were used to screen a Yep24-housed genomic library. A total of 2073 bases of clone Y22 code for a 311 amino acid protein (M(r) 32814), which has similarities to the anion transport proteins: a triplicate gene structure and 6 hydrophobic segments. Typical for PTP, the triplicate gene structure possesses the X-Pro-X-(Asp/Glu)-X-X-(Lys/Arg)-X-(Arg/Lys)-X (X is an unspecified amino acid) motif and the very high homology only between the first and second repeat. The 6 hydrophobic segments harbor most of the 116 amino acids that are conserved between the yeast and the beef proteins. An N-terminal-extended signal sequence, as found in the beef protein, is absent. The yeast protein has about 33% fewer basic and acidic amino acids and five fewer Cys residues than the beef protein. The protein is insensitive to N-ethylmaleimide since Cys-42 (beef) has been replaced with a Thr. Mersalyl sensitivity has been retained and must be due to one of its three cysteines. Among these three cysteines, only Cys-28, located in the first hydrophobic segment, is conserved between the yeast and the beef protein.