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依赖硫化物的光合电子流与蓝藻湖生颤藻类囊体中的质子转运相偶联。

Sulfide-dependent photosynthetic electron flow coupled to proton translocation in thylakoids of the cyanobacterium Oscillatoria limnetica.

作者信息

Shahak Y, Arieli B, Binder B, Padan E

机构信息

Biochemistry Department, Weizmann Institute of Science, Rehovot, Israel.

出版信息

Arch Biochem Biophys. 1987 Dec;259(2):605-15. doi: 10.1016/0003-9861(87)90527-3.

Abstract

Light-induced proton translocation coupled to sulfide-dependent electron transport has been studied in isolated thylakoids of the cyanobacterium Oscillatoria limnetica. The thylakoids are obtained by osmotic shock of washed spheroplasts, prepared with glycine-betaine as the osmotic stabilizer. 13C NMR studies suggests that betaine is the major osmoregulator in O. limnetica. Thylakoid preparations obtained from both sulfide-induced anoxygenic cells and noninduced oxygenic cells are capable of proton pumping coupled to phenazinemethosulfate-mediated cyclic electron flow. However, only in the induced thylakoids can sulfide-dependent proton gradient (delta pH) formation be measured, using either NADP or methyl viologen as the terminal acceptor. Sulfide-dependent delta pH formation correlates with a high-affinity electron donation site (apparent Km 44 microM at pH 7.9). This site is not lost upon washing of the thylakoids. In addition, both sulfide-dependent electron transport and delta pH formation are sensitive to inhibitors of the cytochrome b6f complex such as 2-n-nonyl-4-hydroxyquinoline-N-oxide, 2,4-dinitrophenyl ether of 2-iodo-4-nitrothymol, or stigmatellin. Sulfide-dependent NADP photoreduction of low affinity (which does not saturate by as much as 7 mM sulfide) is detected in both induced and noninduced thylakoids, but this activity is insensitive to the inhibitors and is not coupled to proton transport. It is suggested that the adaptation of O. limnetica to anoxygenic photosynthesis involves the induction of a thylakoid factor(s) which creates a high-affinity site for sulfide, and the transfer of its electrons via the cytochrome b6f complex, coupled to proton translocation.

摘要

在蓝藻湖生颤藻分离的类囊体中,研究了与硫化物依赖性电子传递偶联的光诱导质子转运。类囊体通过用甘氨酸 - 甜菜碱作为渗透稳定剂制备的洗涤过的原生质球的渗透休克获得。13C NMR研究表明,甜菜碱是湖生颤藻中的主要渗透调节剂。从硫化物诱导的无氧细胞和未诱导的有氧细胞获得的类囊体制剂都能够进行与吩嗪硫酸甲酯介导的循环电子流偶联的质子泵浦。然而,只有在诱导的类囊体中,才能使用NADP或甲基紫精作为末端受体来测量硫化物依赖性质子梯度(ΔpH)的形成。硫化物依赖性ΔpH的形成与一个高亲和力电子供体位点相关(在pH 7.9时表观Km为44μM)。该位点在类囊体洗涤后不会丢失。此外,硫化物依赖性电子传递和ΔpH的形成对细胞色素b6f复合物的抑制剂敏感,如2 - n - 壬基 - 4 - 羟基喹啉 - N - 氧化物、2 - 碘 - 4 - 硝基百里酚的2,4 - 二硝基苯基醚或抑藻素。在诱导和未诱导的类囊体中都检测到了低亲和力的硫化物依赖性NADP光还原(即使7 mM硫化物也不能使其饱和),但这种活性对抑制剂不敏感,并且与质子转运不偶联。有人提出,湖生颤藻对无氧光合作用的适应涉及诱导一种类囊体因子,该因子为硫化物创造一个高亲和力位点,并通过细胞色素b6f复合物传递其电子,与质子转运偶联。

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