title: Functional reconstitution of photosystem II with recombinant manganese-stabilizing proteins containing mutations that remove the disulfide bridge.
abstract: The 33-kDa extrinsic subunit of PSII stabilizes the O2-evolving tetranuclear Mn cluster and accelerates O2 evolution. We have used site-directed mutagenesis to replace one or both Cys residues in spinach MSP with Ala. Previous experiments using native and reduced MSP led to the conclusion that a disulfide bridge between these two cysteines is essential both for its binding and its functional properties. We report here that the disulfide bridge, though essential for MSP stability, is otherwise dispensible. The mutation C51A by itself had a delayed effect on MSP function: [C51A]MSP restored normal rates of O2 evolution to PSII but was defective in stabilizing this activity during extended illumination. In contrast, the Cys-free double mutant, [C28A,C51A]MSP, was functionally identical to the wild-type protein. Based on results presented here, we propose a light-dependent interaction between MSP and PSII that occurs only during the redox cycling of the Mn cluster and which is destabilized by the single mutation, C51A.
authors.affiliation: Department of Biology, University of Michigan, Ann Arbor 48109, USA.
authors.forename: S D
authors.initials: SD
authors.lastname: Betts
authors.forename: J R
authors.initials: JR
authors.lastname: Ross
authors.forename: K U
authors.initials: KU
authors.lastname: Hall
authors.forename: E
authors.initials: E
authors.lastname: Pichersky
authors.forename: C F
authors.initials: CF
authors.lastname: Yocum
mesh_terms: D004220:Disulfides / Q000737:chemistry*; D008345:Manganese / Q000494:pharmacology*; D016297:Mutagenesis, Site-Directed; D010100:Oxygen / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D045332:Photosystem II Protein Complex; D011487:Protein Conformation; D011994:Recombinant Proteins; D018724:Spinacia oleracea; D013329:Structure-Activity Relationship
chemical_list: D004220:Disulfides; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D011994:Recombinant Proteins; D008345:Manganese; D010100:Oxygen