Energy-linked anion transport. Cloning, sequencing, and characterization of a full length cDNA encoding the rat liver mitochondrial proton/phosphate symporter.
Energy-linked anion transport. Cloning, sequencing, and characterization of a full length cDNA encoding the rat liver mitochondrial proton/phosphate symporter.
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发表时间：
1989-09
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通讯作者：
Gloria C. Ferreira;R. Pratt;P. Pedersen
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作者：
Gloria C. Ferreira;R. Pratt;P. Pedersen
以牛心脏部分长度磷酸转运蛋白克隆为杂交探针，从文库中获得了编码大鼠肝脏线粒体磷酸转运蛋白前体(H+/PI共同转运体)的全长cDNA克隆。整个克隆全长1263个碱基对，5‘和3’非翻译区分别为16和168个碱基对。开放阅读框编码成熟蛋白(312个氨基酸)，其前有44个富含碱性残基的氨基酸。通过对纯磷转运蛋白的前17个氨基端氨基酸序列的分析，证实了从DNA序列预测的多肽序列。大鼠肝脏磷酸转运蛋白与牛心转运蛋白有32个氨基酸的不同(即大约10%)。它包含一个从139到159的氨基酸区域，该区域与肝脏线粒体ATP合成酶的β亚基有37%的同源性。PI转运体与PI结合蛋白、其他H+连接的转运体以及人葡萄糖转运体的氨基酸序列比较没有发现显著的序列同源性。以大鼠肝线粒体PI载体基因为探针，对大鼠和酿酒酵母基因组DNA进行Southern blotts分析，显示出非常相似的限制性内切酶图谱，这一发现与同源PI载体蛋白在低等和高等真核生物中的存在一致。这是首次报道分离、测序和鉴定编码参与能量偶联PI转运的蛋白质的全长cDNA.
A full length cDNA clone encoding the precursor of the rat liver mitochondrial phosphate transporter (H+/Pi symporter) has been isolated from a cDNA library using a bovine heart partial length phosphate transporter clone as a hybridization probe. The entire clone is 1263 base pairs in length with 5'- and 3'-untranslated regions of 16 and 168 base pairs, respectively. The open reading frame encodes for the mature protein (312 amino acids) preceded by a presequence of 44 amino acids enriched in basic residues. The polypeptide sequence predicted from the DNA sequence was confirmed by analyzing the first 17 amino-terminal amino acids of the pure phosphate transporter protein. The rat liver phosphate transporter differs from the bovine heart transporter in 32 amino acids (i.e. approximately 10%). It contains a region from amino acid 139 to 159 which is 37% identical with the beta-subunit of the liver mitochondrial ATP synthase. Amino acid sequence comparisons of the Pi transporter with Pi binding proteins, other H+-linked symporters, and the human glucose transporter did not reveal significant sequence homology. Analysis of genomic DNA from both rat and S. cerevisiae by Southern blots using the rat liver mitochondrial Pi carrier cDNA as a probe revealed remarkably similar restriction patterns, a finding consistent with the presence in lower and higher eukaryotes of homologous Pi carrier proteins. This is the first report of the isolation, sequencing, and characterization of a full length cDNA coding for a protein involved in energy-coupled Pi transport.