Battchikova Natalia, Zhang Pengpeng, Rudd Stephen, Ogawa Teruo, Aro Eva-Mari
Department of Biology, Plant Physiology and Molecular Biology, FIN-20014 University of Turku, Finland.
J Biol Chem. 2005 Jan 28;280(4):2587-95. doi: 10.1074/jbc.M410914200. Epub 2004 Nov 17.
The subunit compositions of two types of NAD(P)H dehydrogenase complexes of Synechocystis sp. PCC 6803, NDH-1L and NDH-1M, were studied by two-dimensional blue-native/SDS-PAGE followed by electrospray tandem mass spectrometry. Fifteen proteins were observed in NDH-1L including hydrophilic subunits (NdhH, -K, -I, -J, -M, and -N) and hydrophobic subunits (NdhA, -B, -E, -G, -D1, and -F1). In addition, NdhL and a novel subunit, Ssl1690 (designated NdhO), were shown to be components of this complex. All subunits mentioned above were present in the NDH-1M complex except NdhD1 and NdhF1. NdhL and Ssl1690 (NdhO) were homologous to hypothetical proteins encoded by genomic DNA in higher plants, suggesting that chloroplast NDH-1 complexes contain related subunits. Diagnostic sequence motifs were found for both NdhL and NdhO homologous proteins. Analysis of ndhL deletion mutant (M9) revealed the presence of assembled NDH-1L and NDH-1M complexes, but these complexes appear to be functionally impaired in the absence of NdhL. Both NDH-1 complexes were absent in the ndhB deletion mutant (M55).
通过二维蓝色非变性/SDS-PAGE 结合电喷雾串联质谱法,对集胞藻PCC 6803的两种类型的NAD(P)H脱氢酶复合物(NDH-1L和NDH-1M)的亚基组成进行了研究。在NDH-1L中观察到15种蛋白质,包括亲水性亚基(NdhH、-K、-I、-J、-M和-N)和疏水性亚基(NdhA、-B、-E、-G、-D1和-F1)。此外,NdhL和一个新的亚基Ssl1690(命名为NdhO)被证明是该复合物的组成成分。除了NdhD1和NdhF1外,上述所有亚基都存在于NDH-1M复合物中。NdhL和Ssl1690(NdhO)与高等植物基因组DNA编码的假定蛋白质同源,这表明叶绿体NDH-1复合物含有相关亚基。发现了NdhL和NdhO同源蛋白的诊断序列基序。对ndhL缺失突变体(M9)的分析表明存在组装好的NDH-1L和NDH-1M复合物,但在没有NdhL的情况下,这些复合物似乎功能受损。在ndhB缺失突变体(M55)中,两种NDH-1复合物均不存在。