van Haeringen B, Dekker J P, Bloemendal M, Rögner M, van Grondelle R, van Amerongen H
Department of Biophysics, Free University, Amsterdam, The Netherlands.
Biophys J. 1994 Jul;67(1):411-7. doi: 10.1016/S0006-3495(94)80496-5.
We have developed a straightforward method to separate linear-dichroism and birefringence contributions to electric-field induced signals in a conventional birefringence setup. The method requires the measurement of electric birefringence for three different angular positions of the analyzer. It is demonstrated that the presence of linear dichroism can significantly influence the measured signals and lead to completely erroneous calculations of the birefringence signal and field-free decay times if its contribution is not taken into account. The new method is used to determine electric birefringence and linear dichroism of trimeric Photosystem 1 complexes from the cyanobacterium Synechocystis PCC 6803 in the detergents n-dodecyl-beta-D-maltoside and n-octyl-beta-D-glucoside. It is concluded that the orientation of the particles in the field is predominantly caused by a permanent electric dipole moment that is directed parallel to the symmetry axis of the particles. Comparison of the decay times obtained with dodecylmaltoside and octylglucoside supports a model in which the thickness of the disc-like complexes remains similar (7-8 nm) upon replacing dodecylmaltoside by octylglucoside, whereas the diameter increases from 14.4 +/- 0.2 to 16.6 +/- 0.2 nm because of an increased thickness of the detergent layer. This change in diameter is in good agreement with electron-microscopy results on Photosystem 2 complexes in dodecylmaltoside and octylglucoside (Dekker, J. P., E. J. Boekema, H. T. Witt, and M. Rögner. 1988. Biochim. Biophys. Acta 936:307-318). The value of approximately 16.6 nm for the diameter of Photosystem 1 trimers in dodecylmaltoside is in good agreement with recent results obtained from electron microscopy in combination with extensive image analysis (Kruip, J., E. J. Boekema, D. Bald, A. F. Boonstra, and M. Rögner. 1993. J. Biol. Chem. 268:23353-23360).
我们开发了一种简单的方法,用于在传统双折射装置中分离线性二色性和双折射对电场诱导信号的贡献。该方法需要测量分析器在三个不同角度位置的电双折射。结果表明,如果不考虑线性二色性的贡献,其存在会显著影响测量信号,并导致双折射信号和无场衰减时间的计算完全错误。该新方法用于测定来自集胞藻PCC 6803的三聚体光系统I复合物在去污剂正十二烷基-β-D-麦芽糖苷和正辛基-β-D-葡萄糖苷中的电双折射和线性二色性。得出的结论是,颗粒在场中的取向主要由平行于颗粒对称轴的永久电偶极矩引起。用十二烷基麦芽糖苷和辛基葡萄糖苷获得的衰减时间的比较支持了一个模型,即在用辛基葡萄糖苷替代十二烷基麦芽糖苷后,盘状复合物的厚度保持相似(7 - 8纳米),而直径从14.4±0.2纳米增加到16.6±0.2纳米,这是由于去污剂层厚度增加所致。直径的这种变化与在十二烷基麦芽糖苷和辛基葡萄糖苷中光系统II复合物的电子显微镜结果非常一致(德克尔,J.P.,E.J.博克马,H.T.维特,和M.罗格纳。1988年。生物化学与生物物理学报936:307 - 318)。在十二烷基麦芽糖苷中光系统I三聚体的直径约为16.6纳米,这与最近通过电子显微镜结合广泛图像分析获得的结果非常一致(克鲁普,J.,E.J.博克马,D.鲍尔德,A.F.布恩斯特拉,和M.罗格纳。1993年。生物化学杂志268:23353 - 23360)。