Katoh T, Gantt E
Biochim Biophys Acta. 1979 Jun 5;546(3):383-93. doi: 10.1016/0005-2728(79)90075-6.
Photosynthetically active vesicles with attached phycobilisomes from Anabaena variabilis, were isolated and shown to transfer excitation energy from phycobiliproteins to F696 chlorophyll (Photosystem II). The best results were obtained when cells were disrupted in a sucrose/phosphate/citrate mixture (0.3 : 0.5 : 0.3 M, respectively) containing 1.5% serum albumin. The vesicles showed a phycocyanin/chlorophyll ratio essentially identical to that of whole cells, and oxygen evolution rates of 250 mumol O2/h per mg chlorophyll (with 4 mM ferricyanide added as oxidant), whereas whole cells had rates of up to 450. Excitation of the vesicles by 600 nm light produced fluorescence peaks (-196 degrees C) at 644, 662, 685, 695, and 730 nm. On aging of the vesicles, or upon dilution, the fluorescence yield of the 695 nm emission peak gradually decreased with an accompanying increase and final predominant peak at 685 nm. This shift was accompanied by a decrease in the quantum efficiency of Photosystem II activity from an initial 0.05 to as low as 0.01 mol O2/einstein (605 nm), with a lesser change in the Vmax values. The decrease in the quantum efficiency is mainly attributed to excitation uncoupling between phycobilisomes and Photosystem II. It is concluded that the F685 nm emission peak, often exclusively attributed to Photosystem II chlorophyll, arises from more than one component with phycobilisome emission being a major contributor. Vesicles from which phycobilisomes had been removed, as verified by electron microscopy and spectroscopy, had an almost negligible emission at 685 nm.
分离出了附着有藻胆体的可变鱼腥藻光合活性囊泡,并证明其能将激发能从藻胆蛋白转移至F696叶绿素(光系统II)。当细胞在含有1.5%血清白蛋白的蔗糖/磷酸盐/柠檬酸盐混合物(分别为0.3 : 0.5 : 0.3 M)中破碎时,获得了最佳结果。这些囊泡的藻蓝蛋白/叶绿素比值与完整细胞基本相同,每毫克叶绿素的放氧速率为250 μmol O₂/h(添加4 mM铁氰化物作为氧化剂),而完整细胞的放氧速率高达450。用600 nm光激发囊泡,在644、662、685、695和730 nm处产生荧光峰(-196℃)。随着囊泡老化或稀释,695 nm发射峰的荧光产率逐渐降低,同时685 nm处的发射峰增加并最终占主导地位。这种转变伴随着光系统II活性的量子效率从最初的0.05降至低至0.01 mol O₂/爱因斯坦(605 nm),Vmax值变化较小。量子效率的降低主要归因于藻胆体与光系统II之间的激发解偶联。得出的结论是,通常仅归因于光系统II叶绿素的F685 nm发射峰来自多个成分,其中藻胆体发射是主要贡献者。经电子显微镜和光谱学验证,去除了藻胆体的囊泡在685 nm处的发射几乎可以忽略不计。