Loqué D, Lalonde S, Looger L L, von Wirén N, Frommer W B
Carnegie Institution, 260 Panama St, Stanford, California 94305, USA.
Nature. 2007 Mar 8;446(7132):195-8. doi: 10.1038/nature05579. Epub 2007 Feb 11.
Polytopic membrane proteins are essential for cellular uptake and release of nutrients. To prevent toxic accumulation, rapid shut-off mechanisms are required. Here we show that the soluble cytosolic carboxy terminus of an oligomeric ammonium transporter from Arabidopsis thaliana serves as an allosteric regulator essential for function; mutations in the C-terminal domain, conserved between bacteria, fungi and plants, led to loss of transport activity. When co-expressed with intact transporters, mutants inactivated functional subunits, but left their stability unaffected. Co-expression of two inactive transporters, one with a defective pore, the other with an ablated C terminus, reconstituted activity. The crystal structure of an Archaeoglobus fulgidus ammonium transporter (AMT) suggests that the C terminus interacts physically with cytosolic loops of the neighbouring subunit. Phosphorylation of conserved sites in the C terminus are proposed as the cognate control mechanism. Conformational coupling between monomers provides a mechanism for tight regulation, for increasing the dynamic range of sensing and memorizing prior events, and may be a general mechanism for transporter regulation.
多聚体膜蛋白对于细胞摄取和释放营养物质至关重要。为防止有毒物质积累,需要快速关闭机制。在此我们表明,拟南芥寡聚铵转运蛋白的可溶性胞质羧基末端作为功能必需的变构调节剂;细菌、真菌和植物之间保守的C末端结构域中的突变导致转运活性丧失。当与完整的转运蛋白共表达时,突变体使功能亚基失活,但不影响其稳定性。两个无活性转运蛋白的共表达,一个孔有缺陷,另一个C末端缺失,可重建活性。嗜热栖热菌铵转运蛋白(AMT)的晶体结构表明,C末端与相邻亚基的胞质环发生物理相互作用。C末端保守位点的磷酸化被认为是相关的控制机制。单体之间的构象偶联提供了一种紧密调节的机制,用于增加传感和记忆先前事件的动态范围,并且可能是转运蛋白调节的普遍机制。