Kless H, Oren-Shamir M, Malkin S, McIntosh L, Edelman M
Department of Plant Genetics, Weizmann Institute of Science, Rehovot, Israel.
Biochemistry. 1994 Aug 30;33(34):10501-7. doi: 10.1021/bi00200a035.
The region between helices D and E (D-E region) of the D1 protein of photosystem II (PSII) is exposed at the stromal side of the photosynthetic membrane, contains the secondary plastoquinone (QB) binding niche, and is involved in processes at the reducing side of PSII. The role of the D-E region was studied in 27 site-directed mutants generated in the psbAII gene of the cyanobacterium Synechocystis sp. PCC 6803. The photochemical performance of the modified PSII reaction centers was assessed with respect to photoautotrophic growth, oxygen evolution, fluorescence induction, and herbicide inhibition. A few mutations, located at positions presumably involved in essential interactions in the QB binding niche, greatly interfered with PSII performance. On the other hand, mutations in the presumptive loop region between helices D and de resulted in relatively minor effects, indicating a flexible region not critical for photochemical function. Indeed, although more than 80% of the D-E region is phylogenetically invariant, the bulk of the mutations affected the measured parameters only moderately. The significance of the conserved residues appears to be in subtle interactions that optimize the thermodynamic balance between some of the redox components of PSII, as indicated by mild changes in the steady state fluorescence. Many mutations modified tolerances to PSII herbicides. The dispersion of these mutations throughout the D-E region indicates the complex nature of the interactions, direct and indirect, affecting herbicide binding in the QB niche. Mutation of codons Ser221 and Ser222 to Leu221 and Ala222 revealed a new location coordinating the herbicide diuron in the D1 protein.(ABSTRACT TRUNCATED AT 250 WORDS)
光系统II(PSII)的D1蛋白中螺旋D和E之间的区域(D-E区域)暴露于光合膜的基质侧,包含次生质体醌(QB)结合位点,并参与PSII还原侧的过程。在集胞藻属蓝细菌PCC 6803的psbAII基因中产生的27个定点突变体中研究了D-E区域的作用。针对光合自养生长、氧气释放、荧光诱导和除草剂抑制评估了修饰的PSII反应中心的光化学性能。一些位于推测参与QB结合位点中关键相互作用的位置的突变极大地干扰了PSII的性能。另一方面,螺旋D和de之间推测的环区域中的突变产生的影响相对较小,表明该区域具有灵活性,对光化学功能并非至关重要。实际上,尽管D-E区域超过80%在系统发育上是不变的,但大部分突变仅适度影响所测量的参数。如稳态荧光的轻微变化所示,保守残基的重要性似乎在于优化PSII一些氧化还原成分之间热力学平衡的微妙相互作用。许多突变改变了对PSII除草剂的耐受性。这些突变在整个D-E区域的分散表明影响QB位点中除草剂结合的直接和间接相互作用的复杂性。密码子Ser221和Ser222突变为Leu221和Ala222揭示了D1蛋白中一个新的协调除草剂敌草隆的位置。(摘要截短于250字)