title: Mass spectrometry-based footprinting reveals structural dynamics of loop E of the chlorophyll-binding protein CP43 during photosystem II assembly in the cyanobacterium Synechocystis 6803.
abstract: The PSII repair cycle is required for sustainable photosynthesis in oxygenic photosynthetic organisms. In cyanobacteria and higher plants, proteolysis of the precursor D1 protein (pD1) to expose a C-terminal carboxylate group is an essential step leading to coordination of the Mn4CaO5 cluster, the site of water oxidation. Psb27 appears to associate with both pD1- and D1-containing PSII assembly intermediates by closely interacting with CP43. Here, we report that reduced binding affinity between CP43 and Psb27 is triggered by the removal of the C-terminal extension of the pD1 protein. A mass spectrometry-based footprinting strategy was adopted to probe solvent-exposed aspartic and glutamic acid residues on the CP43 protein. By comparing the extent of footprinting between HT3ΔctpAΔ27PSII and HT3ΔctpAPSII, two genetically modified PSII assembly complexes, we found that Psb27 binds to CP43 on the side of Loop E distal to the pseudo-symmetrical D1-D2 axis. By comparing a second pair of PSII assembly complexes, we discovered that Loop E of CP43 undergoes a significant conformational rearrangement due to the removal of the pD1 C-terminal extension, altering the Psb27-CP43 binding interface. The significance of this conformational rearrangement is discussed in the context of recruitment of the PSII lumenal extrinsic proteins and Mn4CaO5 cluster assembly. In addition to CP43's previously known function as one of the core PSII antenna proteins, this work demonstrates that Loop E of CP43 plays an important role in the functional assembly of the Water Oxidizing Center (WOC) during PSII biogenesis.
authors.affiliation: Department of Biology, Washington University, St. Louis, Missouri 63130.
authors.forename: Haijun
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Chemistry, Washington University, St. Louis, Missouri 63130.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Chemistry, Washington University, St. Louis, Missouri 63130.
authors.forename: Richard Y-C
authors.initials: RY
authors.lastname: Huang
authors.affiliation: Department of Biology, Washington University, St. Louis, Missouri 63130; Department of Chemistry, Washington University, St. Louis, Missouri 63130.
authors.forename: Daniel
authors.initials: D
authors.lastname: Weisz
authors.affiliation: Department of Chemistry, Washington University, St. Louis, Missouri 63130.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Gross
authors.affiliation: Department of Biology, Washington University, St. Louis, Missouri 63130. Electronic address: Pakrasi@wustl.edu.
authors.forename: Himadri B
authors.initials: HB
authors.lastname: Pakrasi
mesh_terms: D003432:Cross-Linking Reagents / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D009154:Mutation; D010100:Oxygen / Q000737:chemistry; D010788:Photosynthesis; D045332:Photosystem II Protein Complex / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D013053:Spectrophotometry / Q000379:methods; D046939:Synechocystis / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D045332:Photosystem II Protein Complex; C083100:photosystem II, chlorophyll binding protein, CP-43; D010100:Oxygen
keywords: CP43 Loop E Conformational Rearrangement; Electron Transfer; Mass Spectrometry (MS); Metalloenzymes; PSII Assembly Intermediate; Photosynthesis; Photosystem II; Protein Cross-linking; Protein Footprinting; Psb27
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