Obata S, Nishimura M, Nagai K, Sakihama N, Shin M
Department of Research, Kishida Chemical Co., Ltd., Sanda-shi, Japan.
Arch Biochem Biophys. 1995 Feb 1;316(2):797-802. doi: 10.1006/abbi.1995.1106.
We detected three ferredoxin components, a, b, and c, in green shoots of Japanese radish seedlings by hydrophobic HPLC analysis using a phenyl-5PW column. All components were also present in mature leaves. Component a was further separated into two components, a1 and a2, by reversed-phase HPLC after carboxymethylation. The existence of four ferredoxins in the photosynthetic tissue of a single plant species is reported here for the first time. Each individual ferredoxin component was isolated, and its primary structure was analyzed for comparison with the two leaf ferredoxins, L-Fds A and B, isolated by Wada et al. (J. Biochem. 105, 619-625 (1989)). The primary structures of components a1, a2, and b were all found in 12 possible structures of L-Fd A deducible from its microheterogeneities. This led us to conclude that L-Fd A is a mixture of three ferredoxins corresponding to components a1, a2, and b. The complete primary structure of component c was established by sequencing of the whole protein and of peptides generated by protease digestion. The N-terminal structure of component c was identical with L-Fd B up to the 11th amino acid residue, suggesting that this component is the same ferredoxin as L-Fd B.
我们通过使用苯基 - 5PW 柱的疏水高效液相色谱分析，在日本萝卜幼苗的绿色嫩枝中检测到三种铁氧化还原蛋白成分，即 a、b 和 c。所有成分在成熟叶片中也都存在。成分 a 在羧甲基化后通过反相高效液相色谱进一步分离为两种成分，a1 和 a2。本文首次报道了单一植物物种光合组织中存在四种铁氧化还原蛋白。分离出了每种单独的铁氧化还原蛋白成分，并分析了其一级结构，以便与和田等人（《生物化学杂志》105 卷，619 - 625 页（1989 年））分离出的两种叶片铁氧化还原蛋白 L - Fds A 和 B 进行比较。发现成分 a1、a2 和 b 的一级结构都存在于从 L - Fd A 的微观异质性推导出来的 12 种可能结构中。这使我们得出结论，L - Fd A 是对应于成分 a1、a2 和 b 的三种铁氧化还原蛋白的混合物。通过对整个蛋白质以及蛋白酶消化产生的肽段进行测序，确定了成分 c 的完整一级结构。成分 c 的 N 端结构在第 11 个氨基酸残基之前与 L - Fd B 相同，这表明该成分与 L - Fd B 是同一种铁氧化还原蛋白。
