A light-driven proton pump from Haloterrigena turkmenica:: Functional expression in Escherichia coli membrane and coupling with a H+ co-transporter
A light-driven proton pump from Haloterrigena turkmenica:: Functional expression in Escherichia coli membrane and coupling with a H+ co-transporter
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DOI：
10.1016/j.bbrc.2005.12.181
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发表时间：
2006-03-10
影响因子：
3.1
通讯作者：
Nara, T
中科院分区：
文献类型：
--
作者：
Kamo, N;Hashiba, T;Nara, T
从土耳其盐陆藻（Haloterrigena turkmenica）（JCM 9743）中克隆了编码推定视网膜蛋白的基因。推导的氨基酸序列与δ视紫红质的氨基酸序列最密切相关，δ视紫红质作为光驱动的H+泵发挥作用，并在新菌株Halo terrigena sp. arg-4（K.伊原上村湖Katagiri，T. Kitajima-Ihara，Y. Sugiyama，Y.木村，澳-地Mukohata，古细菌视紫红质的进化：基因复制和功能分化引起的进化速率变化，J. Mol. 285（1999）163-174。GenBank登录号AB 009620）。因此，我们将目前的蛋白质称为H。turkmenica deltarhodopsin（HtdR）在本报告中。与盐生盐杆菌（Halobacteriumsalinarum）的细菌视紫红质（bR）不同，HtdR在大肠杆菌膜中实现了功能性表达，表达量可达10-15 mg蛋白/L培养物。纯化的HtdR的光循环与bR相似。证实了HtdR的光生电质子泵活性。我们在大肠杆菌中共表达了HtdR和EmrE，EmrE是一种质子偶联的多药物外排转运蛋白。大肠杆菌，并成功地挤出乙锭，EmrE的底物，在光照下。(c)2006年爱思唯尔公司All rights reserved.
A gene encoding putative retinal protein was cloned from Haloterrigena turkmenica (JCM9743). The deduced amino acid sequence was most closely related to that of deltarhodopsin, which functions as a light-driven H+ pump and was identified in a novel strain Halo terrigena sp. arg-4 (K. Ihara T. Uemura, l. Katagiri, T. Kitajima-Ihara, Y. Sugiyama, Y. Kimura, Y. Mukohata, Evolution of the archaeal rhodopsins: Evolution rate changes by gene duplication and functional differentiation, J. Mol. Biol. 285 (1999) 163-174. GenBank Accession No. AB009620). Thus, we called the present protein H. turkmenica deltarhodopsin (HtdR) in this report. Differing from the Halobacterium salinarum bacteriorhodopsin (bR), functional expression of HtdR was achieved in Escherichia coli membrane with a high yield of 10-15 mg protein/L Culture. The photocycle of purified HtdR was similar to that of bR. The photo-induced electrogenic proton pumping activity of HtdR was verified. We co-expressed both HtdR and EmrE, a proton-coupled multi-drug efflux transporter in E. coli, and the cells successfully extruded ethidium, a substrate of EmrE, on illumination. (c) 2006 Elsevier Inc. All rights reserved.