The sequence, bacterial expression, and functional reconstitution of the rat mitochondrial dicarboxylate transporter cloned via distant homologs in yeast and Caenorhabditis elegans
The sequence, bacterial expression, and functional reconstitution of the rat mitochondrial dicarboxylate transporter cloned via distant homologs in yeast and Caenorhabditis elegans
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DOI：
10.1074/jbc.273.38.24754
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发表时间：
1998-09-18
影响因子：
4.8
通讯作者：
Walker, JE
中科院分区：
文献类型：
--
作者：
Fiermonte, G;Palmieri, L;Walker, JE
二羧酸载体（DIC）属于线粒体内膜中发现的转运蛋白家族。这。哺乳动物蛋白质的生化特性已被表征，但蛋白质并不丰富。它很难纯化并且尚未测序。我们使用远缘酵母DIC的序列来鉴定秀丽隐杆线虫基因组中编码的相关蛋白，然后，用线虫序列鉴定相关的小鼠表达序列标签，并使用小鼠序列分离大鼠同源物的cDNA，线虫和大鼠蛋白的序列具有线粒体转运蛋白家族的特征。这两种蛋白均在细菌中表达并重构为磷脂囊泡，其运输特性与整个大鼠线粒体和重构为囊泡的大鼠 DIC 的运输特性非常相似。正如 DIC 在糖异生和尿素生成中的作用所预期的那样，它的转录本在大鼠肝脏和肾脏中被检测到，但出乎意料的是，它们也在大鼠心脏和脑组织中被检测到，在这些组织中，该蛋白可能发挥其他作用，可能为克雷布斯循环提供底物。
The dicarboxylate carrier (DIC) belongs to a family of transport proteins found in the inner mitochondrial membranes. The. biochemical properties of the mammalian protein have been characterized, but the protein is not abundant. It is difficult to purify and had not been sequenced. We have used the sequence of the distantly related yeast DIC to identify a related protein encoded in the genome of Caenorhabditis elegans, Then, related murine expressed sequence tags were identified with the worm sequence, and the murine sequence was used to isolate the cDNA for the rat homolog, The sequences of the worm and rat proteins have features characteristic of the family of mitochondrial transport proteins. Both proteins were expressed in bacteria and reconstituted into phospholipid vesicles where their transport characteristics closely resembled those of whole rat mitochondria and of the rat DIC reconstituted into vesicles. As expected from the role of the DIC in gluconeogenesis and ureogenesis, its transcripts were detected in rat liver and kidney, but unexpectedly, they were also detected in rat heart and brain tissues where the protein may fulfill other roles, possibly in supplying substrates to the Krebs cycle.