De Marcos Lousa Carine, Trézéguet Véronique, David Claudine, Postis Vincent, Arnou Bertrand, Pebay-Peyroula Eva, Brandolin Gérard, Lauquin Guy J-M
Laboratoire de Physiologie Moléculaire et Cellulaire, IBGC-CNRS, UMR 5095, 1 rue Camille Saint-Saëns, 33077 Bordeaux Cedex, France.
Biochemistry. 2005 Mar 22;44(11):4342-8. doi: 10.1021/bi0475370.
We isolated yeast Saccharomyces cerevisiae cells transformed with one of the three human adenine nucleotide carrier genes (HANC) that exhibited higher growth capacity than previously observed. The HANC genes were isolated from these clones, and we identified two independent mutations of HANC that led to replacement of valine 181 located in the fourth transmembrane segment by methionine or phenylalanine. Tolerance of this position toward substitution with various amino acids was systematically investigated, and since HANC/V181M was among the most efficient in growth complementation, it was more extensively studied. Here we show that increased growth capacities were associated with higher ADP/ATP exchange activities and not with higher human carrier amount in yeast mitochondria. These results are discussed in the light of the bovine Ancp structure, that shares more than 90% amino acid identity with Hancps, and its interaction with the lipid environment.
我们分离出了用三种人类腺嘌呤核苷酸载体基因（HANC）之一转化的酿酒酵母细胞，这些细胞表现出比先前观察到的更高的生长能力。从这些克隆中分离出HANC基因，我们鉴定出HANC的两个独立突变，这些突变导致位于第四个跨膜段的缬氨酸181被甲硫氨酸或苯丙氨酸取代。系统地研究了该位置对各种氨基酸取代的耐受性，由于HANC/V181M在生长互补方面是最有效的之一，因此对其进行了更广泛的研究。在这里我们表明，生长能力的提高与更高的ADP/ATP交换活性相关，而与酵母线粒体中更高的人类载体量无关。根据与Hancps有超过90%氨基酸同一性的牛Ancp结构及其与脂质环境的相互作用，对这些结果进行了讨论。
