Radiation Biology Laboratory, Smithsonian Institution, 12441 Parklawn Drive, Rockville, Maryland 20852.
Plant Physiol. 1979 Apr;63(4):615-20. doi: 10.1104/pp.63.4.615.
A general procedure for the isolation of functionally intact phycobilisomes was devised, based on modifications of previously used procedures. It has been successful with numerous species of red and blue-green algae (Anabaena variabilis, Anacystis nidulans, Agmenellum quadruplicatum, Fremyella diplosiphon, Glaucosphaera vacuolata, Griffithsia pacifica, Nemalion multifidum, Nostoc sp., Phormidium persicinum, Porphyridium cruentum, P. sordidum, P. aerugineum, Rhodosorus marinus). Isolation was carried out in 0.75 molar K-phosphate (pH 6.8 to 7.0) at 20 to 23 C on sucrose step gradients. Lower temperature (4 to 10 C) was usually unfavorable resulting in uncoupling of energy transfer and partial dissociation of the phycobilisomes, sometimes with complete loss of allophycocyanin. Intact phycobilisomes were characterized by fluorescence emission peaks of 670 to 675 nanometers at room temperature, and 678 to 685 nanometers at liquid nitrogen temperature. Uncoupling and subsequent dissociation of phycobilisomes, in lowered ionic conditions, varied with the species and the degree of dissociation but occurred preferentially between phycocyanin and allophycocyanin, or between phycocyanin and phycoerythrin.
我们设计了一种通用的方法来分离功能完整的藻胆体,该方法基于对先前使用的方法的修改。它已成功应用于许多红藻和蓝藻(变鱼腥藻、鱼腥蓝细菌、四尾栅藻、双鞭甲藻、空球藻、太平洋绿球藻、多歧鱼腥藻、念珠藻、钝顶螺旋藻、紫球藻、铜绿微囊藻、海链藻)。在 20 至 23°C 的 0.75 摩尔 K-磷酸盐(pH6.8 至 7.0)中,在蔗糖分步梯度上进行分离。较低的温度(4 至 10°C)通常不利于能量转移的解偶联和藻胆体的部分解离,有时会导致藻蓝蛋白完全丢失。完整的藻胆体在室温下以 670 至 675 纳米的荧光发射峰和液氮温度下以 678 至 685 纳米的荧光发射峰为特征。在较低的离子条件下,藻胆体的解偶联和随后的解离因物种和解离程度而异,但优先发生在藻蓝蛋白和藻红蛋白之间,或藻蓝蛋白和别藻蓝蛋白之间。