RESUMO
We have recently reported the crystallization of the reaction center of Photosystem II in the presence of detergent mixtures [Adir N (1999) Acta Crystallogr D Biol Crystallogr D55: 891-894]. We have used high performance liquid chromatography, dynamic light scattering, native gel electrophoresis and thermoluminescence measurements to characterize the interaction between these detergent mixtures and RC II, to try and understand their role in the crystallization process. Size exclusion HPLC and dynamic light scattering confirmed that the isolated RC II used for crystallization was exclusively monomeric. Dynamic light scattering measurements show that the detergent mixtures formed single micelles within a limited range of hydrodynamic radii. Both size exclusion HPLC and dynamic light scattering were used to follow the interaction between the detergent mixtures and monomeric RC II. These techniques revealed a decrease in the detergent mixture treated RC II particle size (with respect with the untreated RC II), and that RC II from solubilized crystals contained particles of the same size. Native gel electrophoresis showed that this change in apparent size is not due to the disintegration of the internal structure of the RC II complex. Thermoluminescence measurements of solubilized RC II crystals showed charge recombination from the S(2,3)Q(A) (-) state, indicating that RC II remains functionally viable following detergent mixture treatment and crystallization. The role of the detergent mixtures in the crystallization of RC II is discussed.
