一种参与质子转运、与叶绿体N，N'-二环己基碳二亚胺结合的蛋白脂质的分离。
Isolation of a chloroplast N,N'-dicyclohexylcarbodiimide-binding proteolipid, active in proton translocation.
作者信息
Nelson N, Eytan E, Notsani B E, Sigrist H, Sigrist-Nelson K, Gitler C
出版信息
Proc Natl Acad Sci U S A. 1977 Jun;74(6):2375-8. doi: 10.1073/pnas.74.6.2375.
Abstract
The N,N'-dicyclohexylcarbodiimide-binding proteolipid from lettuce chloroplast membranes has been purified by a novel, rapid technique involving I-butanol extraction and ether precipitation. Reconstitution of this proteolipid into liposomes composed of chloroplast lipids and subsequent incorporation of bacteriorhodopsin resulted in the formation of liposomes exhibiting a light-dependent accumulation of protons. This accumulation was significantly enhanced upon addition of N,N'-dicyclohexylcarbodiimide at concentrations similar to those that inhibit chloroplast adenosinetriphosphatase activity. Radioactively labeled N,N'-dicyclohexylcarbodiimide was found to be incorporated essentially into the proteolipid of the reconstituted liposomes. These results suggest that the functional unit responsible for proton channeling in the chloroplast membrane has been isolated and reconstituted in the native state.
摘要
通过一种涉及异丁醇提取和乙醚沉淀的新颖快速技术，从生菜叶绿体膜中纯化出了N，N'-二环己基碳二亚胺结合蛋白脂质。将这种蛋白脂质重构到由叶绿体脂质组成的脂质体中，随后掺入细菌视紫红质，导致形成了表现出光依赖质子积累的脂质体。当添加浓度与抑制叶绿体腺苷三磷酸酶活性的浓度相似的N，N'-二环己基碳二亚胺时，这种积累显著增强。发现放射性标记的N，N'-二环己基碳二亚胺基本上掺入了重构脂质体的蛋白脂质中。这些结果表明，负责叶绿体膜中质子通道形成的功能单元已被分离并以天然状态重构。
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