title: Manganese-binding proteins of the oxygen-evolving complex.
abstract: The extrinsic 33-kDa protein (P33) was cross-linked covalently to the binding site on P33-depleted PSII preparations which is responsible for reconstitution of photosynthetic water oxidation after PSII preparations have been washed with 1 M CaCl2. Conditions were found in which more than half of the cross-linked protein complexes formed in the PSII preparations retained the ability to catalyze the oxidation of water. The complex is composed of the P33 cross-linked to the D1 and D2 proteins and a 34-kDa protein, which is present in lower abundance than the other three proteins. After solubilization of the membranes with SDS and purification by preparative SDS-PAGE, the complex retains bound manganese and can catalyze the conversion of H2O2 to O2. Calcium and chloride increased the catalase activity of the purified cross-linked complex while lanthanum or hydroxylamine abolished the activity. By use of the specific activity of the H2O2-dependent reaction to follow the extent of purification of the cross-linked complex, the most highly purified complex was determined to contain 0.34 microgram of manganese/180 micrograms of protein. The mole ratio of Mn/protein was calculated to range from 3.6 to 4.5 depending on the assumed stoichiometry of the protein subunits. The results presented here provide direct evidence that one or more of the three proteins that have cross-linked to the P33 are responsible for binding the manganese of the oxygen-evolving complex.
authors.affiliation: Department of Biology, University of Michigan, Ann Arbor 48109-1048.
authors.forename: R
authors.initials: R
authors.lastname: Mei
authors.forename: J P
authors.initials: JP
authors.lastname: Green
authors.forename: R T
authors.initials: RT
authors.lastname: Sayre
authors.forename: W D
authors.initials: WD
authors.lastname: Frasch
mesh_terms: D002374:Catalase / Q000378:metabolism; D002734:Chlorophyll / Q000378:metabolism*; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes; D008345:Manganese / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins; D010940:Plant Proteins / Q000378:metabolism*; D011190:Potassium Cyanide / Q000494:pharmacology; D011485:Protein Binding
chemical_list: D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D010940:Plant Proteins; D002734:Chlorophyll; D008345:Manganese; D002374:Catalase; D011190:Potassium Cyanide; D010100:Oxygen