Shutilova N I, Demidova L N, Kadoshnikova I G, Klimov V V, Zakrzhevskiĭ D A
Biokhimiia. 1982 Feb;47(2):317-22.
Using a previously developed procedure, further optimization of isolation of the functionally active pigment-protein lipid complex of the reaction center of photosystem 2 (PPLC RC PS-2) by DEAE-cellulose chromatography has been carried out. Preliminary chromatographic separation of PPLC RC PS-1 and PS-2 in the presence of a light-harvesting complex has proved to be the most efficient technique. In the latter case the admixture of P700 within PPLC RC PS-2 decreases (1.8-fold on the average) at chromatographic pH value of 8.0. During chromatography at weakly acid, neutral (6.5-7.0) and weakly alkaline (8.3-9.0) values of pH a further, more than 2-fold decrease of P700 admixture is observed. The increase in the degree of separation of PPLC RC PS-1 and PS-2 is concomitant with the increase in PPLC RC PS-2 of photochemically active pheophytin (Ph), an intermediate electron acceptor in RC PS-2 which functions between P680 and Q. The resulting preparations of PPLC RC PS-2 contain one molecule of Ph and 0.06-0.09 molecule of P700 per 31-48 molecules of chlorophyll. The peculiarities of the complexes separation are discussed in terms of the role of hydrogen ion concentration in regulation of affinity between the reaction center complexes and the light-harvesting complex.
采用先前开发的方法,通过DEAE - 纤维素色谱法对光系统2反应中心(PPLC RC PS - 2)的功能活性色素 - 蛋白质脂质复合物的分离进行了进一步优化。在存在捕光复合物的情况下,对PPLC RC PS - 1和PS - 2进行初步色谱分离已被证明是最有效的技术。在后一种情况下,在色谱pH值为8.0时,PPLC RC PS - 2中P700的杂质含量降低(平均降低1.8倍)。在弱酸、中性(6.5 - 7.0)和弱碱性(8.3 - 9.0)pH值下进行色谱分析时,观察到P700杂质含量进一步降低超过2倍。PPLC RC PS - 1和PS - 2分离程度的增加与PPLC RC PS - 2中光化学活性脱镁叶绿素(Ph)的增加相伴,Ph是RC PS - 2中的中间电子受体,在P680和Q之间起作用。所得的PPLC RC PS - 2制剂每31 - 48个叶绿素分子含有一分子Ph和0.06 - 0.09分子P700。根据氢离子浓度在调节反应中心复合物与捕光复合物之间亲和力中的作用,讨论了复合物分离的特点。