van Amerongen H, Kwa S L, van Bolhuis B M, van Grondelle R
Department of Physics and Astronomy, Vrije Universiteit, Amsterdam, The Netherlands.
Biophys J. 1994 Aug;67(2):837-47. doi: 10.1016/S0006-3495(94)80543-0.
Polarized absorption and fluorescence measurements have been performed at 77 K on isotropic and anisotropic preparations of trimeric Light Harvesting Complex II (LHC-II) from spinach. The results enable a decomposition of the absorption spectrum into components parallel and perpendicular to the trimeric plane. For the first time, it is shown quantitatively that the strong absorption band around 676 nm is polarized essentially parallel to the plane of the trimer, i.e., the average angle between the corresponding transition dipole moments and this plane is at most 12 degrees. The different absorption bands for LHC-II should not be considered as corresponding to individual pigments but to collective excitations of different pigments. Nevertheless, the average angle between the Qy transition dipole moments of all chlorophyll a pigments in LHC-II and the trimeric plane could be determined and was found to be 17.5 degrees +/- 2.5 degrees. For the chlorophyll b pigments, this angle is significantly larger (close to 35 degrees). At 77 K, most of the fluorescence stems from a weak band above 676 nm and the corresponding transition dipole moments are oriented further out of plane than the dipole moments corresponding to the 676-nm band. The results are shown to be of crucial significance for understanding the relation between the LHC-II structure and its spectroscopy.
已在77 K下对来自菠菜的三聚体捕光复合物II(LHC-II)的各向同性和各向异性制剂进行了偏振吸收和荧光测量。结果使得能够将吸收光谱分解为与三聚体平面平行和垂直的成分。首次定量显示,676 nm附近的强吸收带基本上与三聚体平面平行偏振,即相应跃迁偶极矩与该平面之间的平均角度最多为12度。LHC-II的不同吸收带不应被视为对应于单个色素,而应被视为不同色素的集体激发。然而,可以确定LHC-II中所有叶绿素a色素的Qy跃迁偶极矩与三聚体平面之间的平均角度,发现其为17.5度±2.5度。对于叶绿素b色素,这个角度明显更大(接近35度)。在77 K时,大部分荧光来自676 nm以上的弱带,并且相应的跃迁偶极矩比对应于676 nm带的偶极矩更偏离平面取向。结果表明对于理解LHC-II结构与其光谱学之间的关系至关重要。