Berry Stephan, Schneider Dirk, Vermaas Wim F J, Rögner Matthias
Lehrstuhl für Biochemie der Pflanzen, Fakultät für Biologie, Ruhr-Universität Bochum, D-44780 Bochum, Germany.
Biochemistry. 2002 Mar 12;41(10):3422-9. doi: 10.1021/bi011683d.
The plastoquinone pool is the central switching point of both respiratory and photosynthetic electron transport in cyanobacteria. Its redox state can be monitored noninvasively in whole cells using chlorophyll fluorescence induction, avoiding possible artifacts associated with thylakoid membrane preparations. This method was applied to cells of Synechocystis sp. PCC 6803 to study respiratory reactions involving the plastoquinone pool. The role of the respiratory oxidases known from the genomic sequence of Synechocystis sp. PCC 6803 was investigated by a combined strategy using inhibitors and deletion strains that lack one or more of these oxidases. The putative quinol oxidase of the cytochrome bd-type was shown to participate in electron transport in thylakoid membranes. The activity of this enzyme in thylakoids was strongly dependent on culture conditions; it was increased under conditions where the activity of the cytochrome b(6)f complex alone may be insufficient for preventing over-reduction of the PQ pool. In contrast, no indication of quinol oxidase activity in thylakoids was found for a second alternative oxidase encoded by the ctaII genes.
质体醌库是蓝细菌中呼吸和光合电子传递的中心转换点。利用叶绿素荧光诱导技术可以在完整细胞中对其氧化还原状态进行非侵入性监测,避免了与类囊体膜制备相关的可能假象。该方法应用于聚球藻属PCC 6803菌株的细胞,以研究涉及质体醌库的呼吸反应。通过使用抑制剂和缺失一种或多种这些氧化酶的缺失菌株的联合策略,研究了从聚球藻属PCC 6803菌株基因组序列中已知的呼吸氧化酶的作用。细胞色素bd型的推定醌氧化酶被证明参与类囊体膜中的电子传递。该酶在类囊体中的活性强烈依赖于培养条件;在仅细胞色素b(6)f复合体的活性可能不足以防止质体醌库过度还原的条件下,其活性会增加。相比之下,未发现由ctaII基因编码的第二种替代氧化酶在类囊体中有醌氧化酶活性的迹象。