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嗜盐嗜碱菌中精氨酸/鸟氨酸反向转运蛋白ArcD的鉴定。

Identification of the arginine/ornithine antiporter ArcD from Halobacterium salinarum.

作者信息

Wimmer Florian, Oberwinkler Tanja, Bisle Birgit, Tittor Jörg, Oesterhelt Dieter

机构信息

Department of Membrane Biochemistry, Max Planck Institut for Biochemistry, Am Klopferspitz 18, D82152 Martinsried, Germany.

出版信息

FEBS Lett. 2008 Nov 12;582(27):3771-5. doi: 10.1016/j.febslet.2008.10.004. Epub 2008 Oct 16.

Abstract

This paper identifies the first arginine/ornithine antiporter ArcD from the domain of archea. The functional role of ArcD is demonstrated by transport assays with radioactive labelled arginine, by its necessity to enable arginine fermentation under anaerobic growth conditions and by the consumption of arginine from the medium during growth. All three experimentally observables are severely disturbed when the deletion strain DeltaArcD is used. The isolated protein is verified by mass spectrometry and reconstituted in vesicles. The proteoliposomes are attached to a membrane and capacitive currents are recorded which appear upon initiation of the transport process by change from arginine-free to arginine-containing buffer. This clearly demonstrates that the purified 34kD protein is the functional unit.

摘要

本文鉴定了来自古菌域的首个精氨酸/鸟氨酸反向转运蛋白ArcD。通过用放射性标记的精氨酸进行转运测定、在厌氧生长条件下实现精氨酸发酵的必要性以及生长过程中从培养基中消耗精氨酸,证明了ArcD的功能作用。当使用缺失菌株DeltaArcD时,所有这三个实验可观察结果都受到严重干扰。通过质谱法验证分离出的蛋白质,并将其重构成囊泡。将蛋白脂质体附着在膜上,并记录电容电流,当从无精氨酸缓冲液变为含精氨酸缓冲液启动转运过程时会出现该电流。这清楚地表明纯化的34kD蛋白是功能单元。

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