The biogenesis of the cyanellae of cyanophora paradoxa. I. Polypeptide composition of the cyanellae
The biogenesis of the cyanellae of cyanophora paradoxa. I. Polypeptide composition of the cyanellae
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蓝藻悖论蓝藻的生物发生。
DOI：
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发表时间：
1989
期刊：
影响因子：
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通讯作者：
R. Trench
中科院分区：
文献类型：
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作者：
R. Burnap;R. Trench
基于使用异源抗体的多肽分离、蛋白质纯化和免疫印迹技术，我们已经能够鉴定出Cyanophora paradoxa蓝藻的几种对光合重要的多肽成分。细胞色素 c-552 和铁氧还蛋白已纯化至电泳均质，并分别表现出 10.5 和 9.0 kDa 的表观分子质量。细胞色素 c-552 的等电点为 pH 4.2±0.1。即使在存在 Cu2+ 的情况下生长的细胞中，质体蓝素在免疫学和光谱上也无法检测到。通过使用 TMBZ 染色程序，已在电泳解析的类囊体样品中定位了细胞色素 f、b-6 和 c-552 的多肽。完整的藻胆体已被纯化并表征了多肽组成以及吸收和发射光谱。光系统 I 和 II 已被分离并表征其光化学活性、光谱特征和多肽组成。光化学活性 PS I 复合物在 77 K 时最大发射波长为 720 nm，包含在变性条件下解析的五个多肽亚基，表观分子量分别为 66、21、18、14 和 11 kDa。 PS II 核心复合物以 140–200 μmol h-1 mg-1 叶绿素的速率介导 1,5-二苯卡巴肼 (DPC) 和 2,6-二氯苯酚靛酚 (DPIP) 之间的光依赖性 3-(3,4-二氯苯基)-1,1-二甲基脲 (DCMU) 敏感电子转移。这些复合物在 77 K 时在 436 和 673 nm 处表现出最大吸收，并在 685 和 695 nm 处表现出荧光发射最大值。通过二维电泳分离 Rubisco 并进行免疫学表征。
Based on polypeptide separation, protein purification and immunoblotting techniques using heterologous antibodies, we have been able to identify several photosynthetically important polypeptide components of the cyanellae of Cyanophora paradoxa. Cytochrome c-552 and ferredoxin have been purified to electrophoretic homogeneity and exhibit apparent molecular masses of 10.5 and 9.0 kDa, respectively. Cytochrome c-552 has an isoelectric point of pH 4.2±0.1. Plastocyanin was immunologically and spectrally undetectable even in cells grown in the presence of Cu2+. Polypeptides for cytochromes f, b-6 and c-552 have been located in electrophoretically resolved thylakoid samples by using the TMBZ-staining procedure. Intact phycobilisomes have been purified and characterized with respect to polypeptide composition and absorption and emission spectra. Photosystems I and II have been isolated and characterized with respect to their photochemical activities, spectral characteristics and polypeptide composition. Photochemically active PS I complexes fluoresce maximally at 720 nm at 77 K and comprise five polypeptide subunits resolved under denaturing conditions with apparent molecular masses of 66, 21, 18, 14 and 11 kDa. PS II core complexes mediate light-dependent 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-sensitive electron transfer between 1,5-diphenylcarbazide (DPC) and 2,6-dichlorophenolindophenol (DPIP) at rates of 140–200 μmol h-1 mg-1 chlorophyll. These complexes exhibit absorption maxima at 436 and 673 nm and show fluorescence emission maxima at 685 and 695 nm at 77 K. Rubisco was separated by two-dimensional electrophoresis and immunologically characterized.