13C ENDOR reveals that the D1 polypeptide C-terminus is directly bound to Mn in the photosystem II oxygen evolving complex.

ABSTRACT: Antiferromagnetically coupled Mn(III)Mn(IV) dimers have been commonly used to study biological systems that exhibit complex exchange interactions. Such is the case for the oxygen evolving complex (OEC) in photosystem II (PSII), where we have studied whether the C-terminal carboxylate of D1-Ala344 is directly bound to the Mn cluster. To probe these protein-derived carboxylate hyperfine interactions, which give direct bonding information, Q-band (34 GHz) Mims ENDOR was performed on a Mn(III)Mn(IV) dimer ([Mn(III)Mn(IV)(mu-O)(2)mu-OAc(TACN)(2)](BPh(4))(2)) (1) that was labeled with (13)C (I = (1)/(2)) at the carboxylate position of the acetate bridge. A(dip) is computed based on atomic coordinates from available X-ray crystal structures to be [-2.4, -0.8, 3.2] MHz. The value for A(iso) was determined based on simulation of the experimental ENDOR data, for complex 1 A(iso) = -1 MHz. Similar studies were then performed on PSII from Synechocystis sp. PCC 6803, in which all alanine-derived C=O groups are labeled with (13)C including the C-terminal alpha-COO(-) group of D1 (Ala344), as well as PSII proteins uniformly labeled with (13)C. Using recent X-ray crystallography data from T. elongatus the values for A(dip) were calculated and simulations of the experimental data led to A(iso) values of 1.2, 1, and 2 MHz, respectively. We infer from complex 1 that an A(iso) significantly larger than 1.2 MHz for a Mn-coordinating carboxylate moiety is unlikely. Therefore, we support the closer arrangement of Ala344 suggested by the Loll and Guskov structures and conclude that the C-terminal carboxylate of D1 polypeptide is directly bound to the Mn cluster.

SUBMITTER: Stull JA 

PROVIDER: S-EPMC3418054 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature