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首页> 外文期刊>Biochemistry >A reconstituted light-harvesting complex from the green sulfur bacterium Chlorobium tepidum containing CsmA and bacteriochlorophyll a
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A reconstituted light-harvesting complex from the green sulfur bacterium Chlorobium tepidum containing CsmA and bacteriochlorophyll a

机译：来自绿色硫细菌绿叶绿藻的重组捕光复合物，含有CsmA和叶绿素a

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摘要

Green sulfur bacteria possess two light-harvesting antenna systems, the chlorosome and the. Fenna-Matthews-Olson (FMO) protein. In addition to self-aggregated bacteriochlorophyll (BChl) c, chlorosomes of Chlorobium tepidum contain a small amount of BChl a (ratio 100:1). The chlorosomal BChl a is associated with CsmA, a 6.2 kDa protein that accounts for more than 50% of the protein content of chlorosomes. This CsmA-BChl a complex is located in the chlorosome baseplate with the hydrophilic C-terminal part of CsmA in contact with the FMO protein. CsmA was purified from Chl. tepidum. Isolated chlorosomes were lyophilized and extracted with chloroform/methanol (1:1, v/v). The extract was further purified using gel filtration and reverse-phase HPLC and the purity of the preparation confirmed by SDS-PAGE. Mass spectrometric analysis showed an m/z of 6154.8, in agreement with the calculated mass of the csmA gene product after C-terminal processing. CD spectroscopy of the isolated protein showed that the main structural motif was an a-helix. We have reconstituted the isolated CsmA protein with BChl a in micelles of n-octyl beta-D-glucopyranoside. The resulting preparation reproduced the spectral characteristics of the CsmA-BChl a complex present in the chlorosome baseplate.
机译：绿硫细菌拥有两个捕光天线系统，即脂质体和。 Fenna-Matthews-Olson（FMO）蛋白。除自聚集的细菌叶绿素（BChl）c外，温带假球藻的含氯体还含有少量的BChla（比例为100：1）。氯体BChla与CsmA相关联，CsmA是6.2 kDa的蛋白质，占脂质体蛋白质含量的50％以上。该CsmA-BChl复合物位于氯体基板中，CsmA的亲水性C端部分与FMO蛋白接触。从Chl纯化CsmA。温热的。将分离的脂质体冻干，并用氯仿/甲醇（1：1，v / v）萃取。使用凝胶过滤和反相HPLC进一步纯化提取物，并通过SDS-PAGE确认制剂的纯度。质谱分析显示m / z为6154.8，与C端处理后csmA基因产物的计算质量相符。分离的蛋白质的CD光谱显示主要结构基序是α-螺旋。我们在n-辛基β-D-吡喃葡萄糖苷的胶束中用BChl a重构了分离的CsmA蛋白。所得制剂再现了CsmA-BChl（一种存在于绿体基质中的复合物）的光谱特征。

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