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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >The yeast mitochondrial ADP/ATP carrier functions as a monomer in mitochondrial membranes
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The yeast mitochondrial ADP/ATP carrier functions as a monomer in mitochondrial membranes

机译：酵母线粒体ADP / ATP载体在线粒体膜中起单体作用

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摘要

Mitochondrial carriers are believed widely to be dimers both in structure and function. However, the structural fold is a barrel of six transmembrane a-helices without an obvious dimerisation interface. Here, we show by negative dominance studies that the yeast mitochondrial ADP/ATP carrier 2 from Saccharomyces cerevisiae (AAC2) is functional as a monomer in the mitochondrial membrane. Adenine nucleotide transport by wild-type AAC2 is inhibited by the sulfhydryl reagent 2-sulfonatoethyl-methanethiosulfonate (MTSES), whereas the activity of a mutant AAC2, devoid of cysteines, is unaffected. Wild-type and cysteine-less AAC2 were coexpressed in different molar ratios in yeast mitochondrial membranes. After addition of MTSES the residual transport activity correlated linearly with the fraction of cysteine-less carrier present in the membranes, and so the two versions functioned independently of each other. Also, the cysteine-less and wild-type carriers were purified separately, mixed in defined ratios and reconstituted into liposomes. Again, the residual transport activity in the presence of MTSES depended linearly on the amount of cysteine-less carrier. Thus, the entire transport cycle for ADP/ATP exchange is carried out by the monomer.
机译：线粒体载体被广泛认为在结构和功能上都是二聚体。但是，结构折叠是一桶六个跨膜的a螺旋，没有明显的二聚化界面。在这里，我们通过负优势研究表明，来自酿酒酵母（AAC2）的酵母线粒体ADP / ATP载体2在线粒体膜中起着单体的作用。巯基试剂2-磺酰基乙基甲烷硫代磺酸盐（MTSES）抑制了野生型AAC2的腺嘌呤核苷酸转运，而没有半胱氨酸的突变AAC2的活性不受影响。野生型和少半胱氨酸的AAC2在酵母线粒体膜中以不同的摩尔比共表达。加入MTSES后，残留的转运活性与膜中存在的无半胱氨酸的载体的比例线性相关，因此这两种形式的功能彼此独立。同样，分别分离纯化无半胱氨酸的载体和野生型载体，以确定的比例混合，然后重新制成脂质体。同样，在存在MTSES的情况下，残留的转运活性与无半胱氨酸载体的量线性相关。因此，用于ADP / ATP交换的整个运输循环由单体进行。

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