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Biochemistry. 2004 Mar 23;43(11):3152-66.

Evidence that the C-terminus of the D1 polypeptide of photosystem II is ligated to the manganese ion that undergoes oxidation during the S1 to S2 transition: an isotope-edited FTIR study.

Author information

1
Department of Biochemistry, University of California, Riverside, California 92521-0129, USA.

Abstract

Isotope-edited FTIR difference spectroscopy was employed to determine if the C-terminal alpha-COO(-) group of the D1 polypeptide ligates the (Mn)(4) cluster in photosystem II (PSII) and, if so, if it ligates the Mn ion that undergoes an oxidation during the S(1) --> S(2) transition. Wild-type and mutant cells of the cyanobacterium Synechocystis sp. PCC 6803 were propagated photoautotrophically in the presence of L-[1-(13)C]alanine or unlabeled ((12)C) L-alanine. In wild-type cells, both the C-terminal alpha-COO(-) group of the D1 polypeptide at D1-Ala344 and all alanine-derived peptide carbonyl groups will be labeled. In D1-A344G and D1-A344S mutant cells, the C-terminal alpha-COO(-) group of the D1 polypeptide will not be labeled because this group is no longer provided by alanine. The resultant S(2)-minus-S(1) FTIR difference spectra of purified wild-type and mutant PSII particles showed that one symmetric carboxylate stretching mode that is altered during the S(1) --> S(2) transition is sensitive to L-[1-(13)C]alanine-labeling in wild-type PSII particles but not in D1-A344G and D1-A344S PSII particles. Because the only carboxylate group that can be labeled in the wild-type PSII particles but not in the mutant PSII particles is the C-terminal alpha-COO(-) group of the D1 polypeptide, we assign the L-[1-(13)C]alanine-sensitive symmetric carboxylate stretching mode to the alpha-COO(-) group of D1-Ala344. In unlabeled wild-type PSII particles, this mode appears at approximately 1356 cm(-1) in the S(1) state and at approximately 1339 or approximately 1320 cm(-1) in the S(2) state. These frequencies are consistent with unidentate ligation of the (Mn)(4) cluster by the alpha-COO(-) group of D1-Ala344 in both the S(1) and S(2) states. The apparent 17-36 cm(-1) downshift in frequency in response to the S(1) --> S(2) transition is consistent with the alpha-COO(-) group of D1-Ala344 ligating a Mn ion whose charge increases during the S(1) --> S(2) transition. Accordingly, we propose that the alpha-COO(-) group of D1-Ala344 ligates the Mn ion that undergoes an oxidation during the S(1) --> S(2) transition. Control experiments were conducted with Mn-depleted wild-type PSII particles. These experiments showed that tyrosine Y(D) may be structurally coupled to the carbonyl oxygen of an alanine-derived peptide carbonyl group.

PMID:
15023066
DOI:
10.1021/bi035915f
[Indexed for MEDLINE]

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