层理鞭枝藻藻红蓝蛋白α亚基的分离纯化和结晶  

PURIFICATION AND CRYSTALLIZATION OF PHYCOERYTHROCYANIN α-SUBUNIT FROM MASTIGOCLADUS LAMINOSUS

作　　者：张震宇[1] 周明[2] 赵开弘[2] 张季平 常文瑞 梁栋材 

机构地区：[1]武汉大学生命科学学院 [2]华中科技大学生命科学与技术学院,湖北武汉430074 [3]中科院生物物理所生物大分子国家重点实验室,北京100101

出　　处：《生物物理学报》2000年第4期667-672,共6页Acta Biophysica Sinica

基　　金：国家自然科学基金!(项目编号 :39770175)资助

摘　　要：藻红蓝蛋白α -亚基具有高效可逆光致变色性质。采用DEAE -纤维素柱层析、SephadexG -75凝胶过滤、FPLC -Superdex75HR10/30柱层析等纯化方法 ,得到了电泳纯的藻红蓝蛋白α-亚基 ,光化学活性达93%。在室温、避光条件下 ,采用悬滴汽相扩散法 ,得到了两种外形不同的藻红蓝蛋白α -亚基晶体 ,进一步证实了上述纯化方法的可靠性。Phycoerythrocyanin is an integral component of phycobilisome in several species of cyanobacteria. Unlike other phycobiliproteins, isolated phycoerythrocyanin α-subunit shows a remarkable reversible photochromism. The classic photochromism of phycoerythrocyanin α-subunit involves the reversible shift of the visible absorption maximum when they are irradiated with light of certain wavelength. Its basis is the structure variation of the chromophore and the apoprotein. Phycoerythrocyanin α-subunit was purified to study the mechanism of its reversible photochromism through X-ray crystallographic studies. Mastigocladus laminosus was collected and disrupted by ultra sonification. The phycobiliproteins were precipitated by stepwise salting out with ammonium sulfate between 30% and 70% saturation. After desalting, the phycobiliproteins were purified by DEAE-cellulose chromatography. Phycoerythrocyanin α-subunit was isolated by Sephadex G-75 column eluted with 63 mmol/L formic acid. A further purification by FPLC can remove degradation peptides of phycoerythrocyanin α-subunit. Photoactivity of final purified phycoerythrocyanin α-subunit was 93%. Screening various conditions, two forms of crystals of phycoerythrocyanin α-subunit were obtained.

关 键 词：藻红蓝蛋白α-亚基 纯化 结晶 分离 

分 类 号：Q946.1[生物学—植物学] Q949.2