Mamedov F, Stefansson H, Albertsson P A, Styring S
Department of Biochemistry, Center for Chemistry and Chemical Engineering, P.O. Box 124, Lund University, S-221 00 Lund, Sweden.
Biochemistry. 2000 Aug 29;39(34):10478-86. doi: 10.1021/bi992877k.
The electron transport properties of photosystem II (PSII) from five different domains of the thylakoid membrane were analyzed by flash-induced fluorescence kinetics. These domains are the entire grana, the grana core, the margins from the grana, the stroma lamellae, and the Y100 fraction (which represent more purified stroma lamellae). The two first fractions originate from appressed grana membranes and have PSII with a high proportion of O(2)-evolving centers (80-90%) and efficient electron transport on the acceptor side. About 30% of the granal PSII centers were found in the margin fraction. Two-thirds of those PSII centers evolve O(2), but the electron transfer on the acceptor side is slowed. PSII from the stroma lamellae was less active. The fraction containing the entire stroma has only 43% O(2)-evolving PSII centers and slow electron transfer on the acceptor side. In contrast, PSII centers of the Y100 fraction show no O(2) evolution and were unable to reduce Q(B). Flash-induced fluorescence decay measurements in the presence of DCMU give information about the integrity of the donor side of PSII. We were able to distinguish between PSII centers with a functional Mn cluster and without any Mn cluster, and PSII centers which undergo photoactivation and have a partially assembled Mn cluster. From this analysis, we propose the existence of a PSII activity gradient in the thylakoid membrane. The gradient is directed from the stroma lamellae, where the Mn cluster is absent or inactive, via the margins where photoactivation accelerates, to the grana core domain where PSII is fully photoactivated. The photoactivation process correlates to the PSII diffusion along the membrane and is initiated in the stroma lamellae while the final steps take place in the appressed regions of the grana core. The margin domain is seemingly very important in this process.
通过闪光诱导荧光动力学分析了类囊体膜五个不同区域的光系统II(PSII)的电子传递特性。这些区域是整个基粒、基粒核心、基粒边缘、基质类囊体和Y100组分(代表更纯化的基质类囊体)。前两个组分起源于紧密堆积的基粒膜,其PSII具有高比例的放氧中心(80 - 90%),并且在受体侧具有高效的电子传递。约30%的基粒PSII中心存在于边缘组分中。其中三分之二的PSII中心能放氧,但受体侧的电子传递减慢。基质类囊体中的PSII活性较低。包含整个基质的组分只有43%的放氧PSII中心,且受体侧的电子传递缓慢。相比之下,Y100组分的PSII中心不放氧,并且无法还原Q(B)。在存在敌草隆(DCMU)的情况下进行的闪光诱导荧光衰减测量提供了有关PSII供体侧完整性的信息。我们能够区分具有功能性锰簇和没有任何锰簇的PSII中心,以及经历光激活并具有部分组装锰簇的PSII中心。通过该分析,我们提出在类囊体膜中存在PSII活性梯度。该梯度从不存在或无活性锰簇的基质类囊体开始,经光激活加速的边缘区域,指向PSII完全光激活的基粒核心区域。光激活过程与PSII沿膜的扩散相关,在基质类囊体中起始,而最终步骤发生在基粒核心的紧密堆积区域。边缘区域在这个过程中似乎非常重要。