Storch K F, Rudolph J, Oesterhelt D
Max-Planck Institut für Biochemie, D-82152 Martinsried, Germany.
EMBO J. 1999 Mar 1;18(5):1146-58. doi: 10.1093/emboj/18.5.1146.
A new metabolic signaling pathway for arginine, both a chemoeffector and a fermentative energy source, is described for Halobacterium salinarum. Systematic screening of 80+ potentially chemotactic compounds with two behavioral assays identified leucine, isoleucine, valine, methionine, cysteine, arginine and several peptides as strong chemoattractants. Deletion analysis of a number of potential halobacterial transducer genes led to the identification of Car, a specific cytoplasmic arginine transducer which lacks transmembrane helices and was biochemically shown to be localized in the cytoplasm. Flow assays were used to show specific adaptive responses to arginine and ornithine in wild-type but not Deltacar cells, demonstrating the role of Car in sensing arginine. The signaling pathway from external arginine to the flagellar motor of the cell involves an arginine:ornithine antiporter which was quantitatively characterized for its transport kinetics and inhibitors. By compiling the chemotactic behavior, the adaptive responses and the characteristics of the arginine:ornithine antiporter to arginine and its analogs, we now understand how the combination of arginine uptake and its metabolic conversion is required to build an effective sensing system. In both bacteria and the archaea this is the first chemoeffector molecule of a soluble methylatable transducer to be identified.
本文描述了嗜盐嗜盐菌中精氨酸的一种新的代谢信号通路,精氨酸既是一种化学效应物,也是一种发酵能量来源。通过两种行为分析方法对80多种潜在趋化性化合物进行系统筛选,确定亮氨酸、异亮氨酸、缬氨酸、蛋氨酸、半胱氨酸、精氨酸和几种肽为强趋化剂。对多个潜在的嗜盐菌转导基因进行缺失分析,鉴定出Car,一种特定的细胞质精氨酸转导蛋白,它缺乏跨膜螺旋,生化分析表明其定位于细胞质中。流式分析用于显示野生型细胞而非缺失Car基因的细胞对精氨酸和鸟氨酸的特异性适应性反应,证明了Car在感知精氨酸中的作用。从细胞外精氨酸到细胞鞭毛马达的信号通路涉及一种精氨酸:鸟氨酸反向转运蛋白,对其转运动力学和抑制剂进行了定量表征。通过汇总趋化行为、适应性反应以及精氨酸:鸟氨酸反向转运蛋白对精氨酸及其类似物的特性,我们现在了解了精氨酸摄取与其代谢转化的组合如何构建一个有效的传感系统。在细菌和古细菌中,这是首次鉴定出的可溶性可甲基化转导蛋白的化学效应分子。