Deng Yong, Ye Ji-Yu, Mi Hua-Ling, Shen Yun-Gang
State Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200032, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Aug;35(8):723-7.
Many efforts have been paid to the separation of an integrated NA(D)PH dehydrogenase (NDH) complex. Several hydrophilic subcomplexes of NDH have been purified from the cyanobacterium Synechocystis PCC6803. However, no hydrophobic NDH subcomplex has ever been separated from cyanobacteria yet. In this paper, two NDH subcomplexes were separated from n-dodecyl beta-D-maltoside(DM)-treated whole cell extracts of Synechocystis PCC6803 by anion exchange chromatography and gel filtration. Both subcomplexes contained the hydrophobic subunit NdhA, suggesting that they were hydrophobic NDH subcomplexes. Of the two subcomplexes, only one subcomplex contained NdhH. These subcomplexes showed NADPH-nitroblue tetrazolium (NBT) oxidoreductase activity and could specifically oxidize NADPH when several quinone analogues were used as electron acceptors, such as ferricyanide, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), 2,6-dichlorophenol indophenol (DCPIP), duroquinone, ubiquinone-0 (UQ-0), etc.
人们为分离完整的NA(D)PH脱氢酶（NDH）复合物付出了诸多努力。已从集胞藻PCC6803中纯化出了几种亲水性的NDH亚复合物。然而，尚未从蓝细菌中分离出疏水性的NDH亚复合物。在本文中，通过阴离子交换色谱法和凝胶过滤，从经正十二烷基-β-D-麦芽糖苷（DM）处理的集胞藻PCC6803全细胞提取物中分离出了两种NDH亚复合物。两种亚复合物均含有疏水性亚基NdhA，这表明它们是疏水性的NDH亚复合物。在这两种亚复合物中，只有一种含有NdhH。当使用几种醌类似物作为电子受体时，如铁氰化物、2,5-二溴-3-甲基-6-异丙基-对苯醌（DBMIB）、2,6-二氯酚靛酚（DCPIP）、杜醌、泛醌-0（UQ-0）等，这些亚复合物表现出NADPH-硝基蓝四唑（NBT）氧化还原酶活性，并且能够特异性地氧化NADPH。
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