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假根羽藻Cyt b6f蛋白复合体的分离与纯化
  • ISSN号:0029-814X
  • 期刊名称:《海洋与湖沼》
  • 时间:0
  • 分类:Q5[生物学—生物化学]
  • 作者机构:[1]中国科学院植物研究所光合作用与环境分子生理学重点实验室,北京100093, [2]中国科学院海洋研究所实验海洋生物学重点实验室,青岛266071
  • 相关基金:国家自然科学基金资助项目,39890390号和30370347号.
中文摘要:

改进了以往分离纯化Cyt b6f的方法,进行了海洋绿藻——假根羽藻(Bryopsis corticulans)Cyt b6f蛋白复合体分离纯的研究。结果表明,该Cyt b6f制剂由Cytf、Cyt b6、Rieske[Fes]蛋白和亚基IV四种主要的多肽亚基以及少量的Chl.a和类胡萝卜素组成。4个多肽亚基的表观分子量分别为34.8、24.0、18.7及16.7kD,该Cyt b6f制剂的Cyt b6/f比值接近2.0,其纯度值为9.9nmolCytf/mg,其催化电子传递的活性(C10-PQH2→PC)为73e/s。上述分析表明,假根羽藻Cyt b6f在组成、纯度和催化活性方面均与已报道的高等植物、淡水绿藻和光合细菌的Cyt b6f制剂相似。

英文摘要:

As a plastoquinol-plastocyanin oxidoreductase, cytochrome b6f complex plays an important role in electron transfer and energy transduction in photosynthesis. It mediates the linear electron flow between PSII and PSI, catalyzes the cyclic electron flow around PSI, and sets up a transmembrane proton electrochemical potential to support energy to form ATP. In addition, cytochrome b6f complex is also involved in the regulation of balanced light excitation energy distribution between photosystems since its redox states governs the activation of LHCII kinase. It consists of four major subunits( Cyt f, Cyt b, Rieske iron-sulfur protein, and subunit IV) and four small subunits(pet G, L, M, and N). Besides, about one chlorophyll a molecule and one or less than one carotenoid molecule have been found in each cytochrome b6f monomer. The cytochrome b6f complex has been purified from higher plants, freshwater green algae and cyanobacteria. However, no case has been reported of this complex from marine green alga. In this work, the cytochrome b6f complex was purified from a marine green alga Bryopsis corticulans successfully by an improved method, its composition and activity were studied in this paper. According to the purification of spinach b6f complex, Cyt b6f was not isolated from Bryopsis corticulans thylakoid. It was shown that most of the b6f complex has gone before last ammonium sulfate fractionations. To solve this problem, ammonium sulfate concentration was adjusted. Although this modification was proved to be useful, some 40-60kD extrinsic proteins still existed. To effectively remove these extrinsic proteins, 2mol/L NaBr membrane washing was repeated after that the extrinsic proteins were removed successfully shown by SDS-PAGE. This modified purification procedures for Bryopsis corticulans b6f complex are:Chloroplasts were isolated from 2kg pre-chilled fresh Bryopsis corticulans as regular way. After osmotically broken, the chloroplasts were resuspended in 2mol/L NaBr, 10mmol/L Tricine-NaOH(pH 8.0?

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期刊信息
  • 《海洋与湖沼》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中国海洋湖沼学会
  • 主编:孙松
  • 地址:青岛市南海路7号中科院海洋所内
  • 邮编:266071
  • 邮箱:
  • 电话:0532-82898753
  • 国际标准刊号:ISSN:0029-814X
  • 国内统一刊号:ISSN:37-1149/P
  • 邮发代号:2-421
  • 获奖情况:
  • 2000年中国科学院优秀期刊一等奖,2000年山东省科委、新闻出版黑署评估优秀级,中国期刊方阵“双百”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:21021