Crystallization of phycoerythrin 545 of Rhodomonas lens using detergents and unusual additives

被引:15
作者
Becker, M [1 ]
Stubbs, MT [1 ]
Huber, R [1 ]
机构
[1] Max Planck Inst Biochem, Abt Strukturforsch, D-82152 Martinsried, Germany
关键词
Cryptophyceae; detergent; light-harvesting protein; photosynthesis; phycobiliprotein; propionamide; protein crystal; X-ray diffraction;
D O I
10.1002/pro.5560070306
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Phycoerythrin 545 from the cryptomonad alga, Rhodomonas lens, has been crystallized under a wide variety of conditions. Although this type of photosynthetic light-harvesting protein is water soluble, detergents were always required for crystallization. The crystals were typically poorly ordered, or ordered in only two dimensions. However, crystals that were well-ordered in three dimensions could be obtained under two different conditions. Both used polyethylene glycol as precipitant and the detergent lauryldimethylaminoxide, but the additives that were critical for obtaining well-ordered crystals were propionamide in one case and Cs+/Br- in the other. Crystals obtained in the presence of propionamide have the space group P2(1)2(1)2(1), with cell constants of a = 85.6 Angstrom, b = 108.2 Angstrom, and c = 131.0 Angstrom, and contain two dimers [i.e., 2 X (alpha(2) beta(2))] in the asymmetric unit. They show diffraction to at least 3.0 Angstrom resolution. The crystals grown with Cs+/Br- are nearly isomorphous. Both types of crystals show intense, strongly polarized fluorescence, suggesting that energy transfer in the crystals is highly efficient. This should provide a basis for quantitative investigation of the role of exciton interactions in energy transfer in cryptomonad phycobiliproteins.
引用
收藏
页码:580 / 586
页数:7
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