title: Electric field effects on the chlorophylls, pheophytins, and beta-carotenes in the reaction center of photosystem II.
abstract: We present an electric field modulated absorption spectroscopy (Stark effect) study of isolated photosystem II reaction center complexes, including a preparation in which the inactive pheophytin H(B) was exchanged for 13(1)-deoxo-13(1)-hydroxy-pheophytin. The results reveal that the Stark spectrum of the Q(x) and Q(y) transitions of the pheophytins has a second-derivative line shape, indicating that the Stark effect is dominated by differences in the dipole moment between the ground and the electronically excited states of these transitions (Delta mu). The Delta mu values for the Q(x) and Q(y) transitions of H(B) are small (Delta mu = 0.6-1.0 D f(-1)), whereas that of the Q(x) transition of the active pheophytin H(A) is remarkably large (Delta mu = 3 D f(-1)). The Stark spectrum of the red-most absorbing pigments also shows a second-derivative line shape, but this spectrum is considerably red-shifted as compared to the second derivative of the absorption spectrum. This situation is unusual but has been observed before in heterodimer special pair mutants of purple bacterial reaction centers [Moore, L. J., Zhou, H., and Boxer, S. G. (1999) Biochemistry 38, 11949-11960]. The red-shifted Stark spectra can be explained by a mixing of exciton states with a charge-transfer state of about equal energy. We conclude that the charge transfer state involves H(A) and its immediate chlorophyll neighbor (B(A)), and we suggest that this (B(A)(delta+)H(A)(delta-)) charge transfer state plays a crucial role in the primary charge separation reaction in photosystem II. In contrast to most other carotenes, the two beta-carotene molecules of the photosystem II reaction center display a very small Delta mu, which can most easily be explained by excitonic coupling of both molecules. These results favor a model that locates both beta-carotene molecules at the same side of the complex.
authors.affiliation: Division of Physics and Astronomy, Faculty of Sciences, Vrije Universiteit, De Boelelaan 1081, 1081 HV Amsterdam, The Netherlands.
authors.forename: Raoul N
authors.initials: RN
authors.lastname: Frese
authors.forename: Marta
authors.initials: M
authors.lastname: Germano
authors.forename: Frank L
authors.initials: FL
authors.lastname: de Weerd
authors.forename: Ivo H M
authors.initials: IH
authors.lastname: van Stokkum
authors.forename: Anatoli Ya
authors.initials: AY
authors.lastname: Shkuropatov
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Shuvalov
authors.forename: Hans J
authors.initials: HJ
authors.lastname: van Gorkom
authors.forename: Rienk
authors.initials: R
authors.lastname: van Grondelle
authors.forename: Jan P
authors.initials: JP
authors.lastname: Dekker
mesh_terms: D002734:Chlorophyll / Q000737:chemistry*; D004574:Electromagnetic Fields; D045342:Light-Harvesting Protein Complexes; D010674:Pheophytins / Q000737:chemistry*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045332:Photosystem II Protein Complex; D013057:Spectrum Analysis / Q000379:methods; D018724:Spinacia oleracea; D019207:beta Carotene / Q000737:chemistry*
chemical_list: D045342:Light-Harvesting Protein Complexes; D010674:Pheophytins; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D019207:beta Carotene; C070970:photosystem II, chlorophyll-binding protein, CP-47; D002734:Chlorophyll