Lind L K, Shukla V K, Nyhus K J, Pakrasi H B
Department of Biology, Washington University, St. Louis, Missouri 63130-4899.
J Biol Chem. 1993 Jan 25;268(3):1575-9.
The psbJ gene is a member of the psbEFLJ gene cluster in the cyanobacterium Synechocystis sp. PCC 6803 as well as in the chloroplasts of green plants. The putative product of the psbJ gene is a 4-kDa protein with one membrane-spanning domain. We have raised rabbit antibodies against a T7 gene 10-psbJ fusion protein, overexpressed in Escherichia coli. These antibodies recognized a polypeptide of expected size in the thylakoid membrane from wild type Synechocystis cells. We have also created a targeted mutant of Synechocystis 6803 in which the fourth codon of the psbJ open reading frame was modified to a translational stop codon. Thylakoid membranes from these mutant cells lacked the protein recognized by the antibodies. In the mutant cells, the partial electron transfer reaction mediated by the photosystem I complex was unaffected, whereas the rate of the photosystem II (PSII)-mediated reaction was 46% of that in wild type cells. Herbicide binding assays indicated that the PSII to chlorophyll ratio in the mutant cells was 49% of that in wild type cells. These results indicate that while the PsbJ protein is not essential for the photochemical activity it controls the amount of functionally assembled PSII complex in the thylakoid membrane.
psbJ基因是蓝藻集胞藻PCC 6803以及绿色植物叶绿体中psbEFLJ基因簇的成员之一。psbJ基因的推定产物是一种具有一个跨膜结构域的4 kDa蛋白质。我们制备了针对在大肠杆菌中过表达的T7基因10 - psbJ融合蛋白的兔抗体。这些抗体识别野生型集胞藻细胞类囊体膜中预期大小的多肽。我们还构建了集胞藻6803的靶向突变体,其中psbJ开放阅读框的第四个密码子被修饰为翻译终止密码子。这些突变体细胞的类囊体膜缺乏抗体识别的蛋白质。在突变体细胞中,由光系统I复合物介导的部分电子传递反应未受影响,而光系统II(PSII)介导的反应速率是野生型细胞的46%。除草剂结合试验表明,突变体细胞中PSII与叶绿素的比率是野生型细胞的49%。这些结果表明,虽然PsbJ蛋白对于光化学活性不是必需的,但它控制着类囊体膜中功能组装的PSII复合物的数量。