From the Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology and.
J Biol Chem. 2014 May 9;289(19):13374-84. doi: 10.1074/jbc.M114.547448. Epub 2014 Mar 20.
The human genome encodes 53 members of the solute carrier family 25 (SLC25), also called the mitochondrial carrier family, many of which have been shown to transport carboxylates, amino acids, nucleotides, and cofactors across the inner mitochondrial membrane, thereby connecting cytosolic and matrix functions. In this work, a member of this family, SLC25A29, previously reported to be a mitochondrial carnitine/acylcarnitine- or ornithine-like carrier, has been thoroughly characterized biochemically. The SLC25A29 gene was overexpressed in Escherichia coli, and the gene product was purified and reconstituted in phospholipid vesicles. Its transport properties and kinetic parameters demonstrate that SLC25A29 transports arginine, lysine, homoarginine, methylarginine and, to a much lesser extent, ornithine and histidine. Carnitine and acylcarnitines were not transported by SLC25A29. This carrier catalyzed substantial uniport besides a counter-exchange transport, exhibited a high transport affinity for arginine and lysine, and was saturable and inhibited by mercurial compounds and other inhibitors of mitochondrial carriers to various degrees. The main physiological role of SLC25A29 is to import basic amino acids into mitochondria for mitochondrial protein synthesis and amino acid degradation.
人类基因组编码了 53 种溶质载体家族 25(SLC25)成员,也称为线粒体载体家族,其中许多已被证明可在内膜上转运羧酸、氨基酸、核苷酸和辅因子,从而连接细胞质和基质功能。在这项工作中,该家族的一个成员 SLC25A29,先前被报道为一种线粒体肉碱/酰基辅酶 A 或鸟氨酸样载体,已在生化上进行了彻底的表征。SLC25A29 基因在大肠杆菌中过表达,基因产物在磷脂囊泡中被纯化和重建。其转运特性和动力学参数表明,SLC25A29 转运精氨酸、赖氨酸、同型精氨酸、甲基精氨酸,以及程度较小的鸟氨酸和组氨酸。肉碱和酰基辅酶 A 不能被 SLC25A29 转运。该载体除了反向交换转运外,还催化大量的单向转运,对精氨酸和赖氨酸表现出高转运亲和力,并对汞化合物和其他线粒体载体抑制剂有不同程度的饱和和抑制作用。SLC25A29 的主要生理作用是将碱性氨基酸导入线粒体,用于线粒体蛋白质合成和氨基酸降解。