Chemically modified reaction centers of photosystem II: Exchange of pheophytin a with 7-deformyl-7-hydroxymethyl-pheophytin b

• Published in: Biochimica et biophysica acta 2014-11, Vol.1837 (11), p.1870-1881
• Main Authors: Zabelin, Alexey A., Shkuropatova, Valentina A., Makhneva, Zoya K., Moskalenko, Andrey A., Shuvalov, Vladimir A., Shkuropatov, Anatoly Ya
• Format: Article
• Language: English
• Subjects: 7-Deformyl-7-hydroxymethyl-pheophytin b; Charge separation; Chemical exchange; Pheophytin a; Photosystem II; Reaction center
• ISSN: 0005-2728; 0006-3002; 1879-2650
• DOI: 10.1016/j.bbabio.2014.08.004

The native pheophytin a (Pheo a) in isolated reaction centers of photosystem II (PSII RCs) has been chemically exchanged with extraneous 7-deformyl-7-hydroxymethyl-Pheo b (71-OH-Pheo b) which differs from Pheo a by the C-7 substituent (hydroxymethyl instead of methyl). The two pigments have similar reduction potentials in vitro [M. Meyer, Dissertation, Universität München, 1997], while their absorption spectra show small but distinct differences in the visible region. The resulting 71-OH-Pheo b-modified reaction center preparations were characterized by high-performance liquid chromatography, electronic absorption and light-induced Fourier transform infra red absorption difference spectroscopies, together with photoaccumulation of the reduced pheophytin electron acceptor and NaBH4-treatment. About 70% of the total Pheo a molecules are found to be replaced by 71-OH-Pheo b molecules in modified preparations, indicating that both the photochemically active (PheoD1) and inactive (PheoD2) binding sites were subjected to pigment exchange. The 71-OH-Pheo b molecule located at the PheoD1 site is able to functionally replace the native Pheo a, participating in primary charge separation as an electron acceptor. The Qx absorption band of this modified pheophytin molecule is localized at ~546nm; its Qy band is blue-shifted with respect to the absorption of other reaction center core pigments, being located at ~665nm. The Qy and Qx optical transitions of the 71-OH-Pheo b molecule exchanged into the PheoD2 site are identified at 677 and 543.5nm, respectively. The photochemically active double-modified PSII RCs additionally containing 7-deformyl-7-hydroxymethyl-131-deoxo-131-hydroxy-Pheo b at the PheoD2 site were obtained by treatment of the 71-OH-Pheo b-modified RCs with NaBH4. [Display omitted] •The native Pheo a in PSII RCs has been chemically exchanged with 7-deformyl-7-hydroxymethyl-Pheo b (71-OH-Pheo b).•Both PheoD1 and PheoD2 binding sites were subjected to pigment exchange.•The 71-OH-Pheo b molecule at the PheoD1 site is able to participate in primary charge separation as an electron acceptor.•The Qy and Qx optical transitions of 71-OH-Pheo b at PheoD1,D2 sites were identified.