title: In vivo and in vitro photoinhibition reactions generate similar degradation fragments of D1 and D2 photosystem-II reaction-centre proteins.
abstract: Isolation of photosystem-II reaction centres from pea leaves after photoinhibitory treatment at low temperature (0-1 degrees C) has provided evidence for the mechanism of degradation of the D1 protein in vivo. These isolated reaction centres did not appear to be spectrally distinct from preparations obtained from control leaves that had not been photoinhibited. Breakdown fragments of both the D1 and D2 proteins were, however, found in preparations isolated from photoinhibited leaves, and showed similarities with those detected when isolated reaction centres were exposed to acceptor-side photoinhibition. Analyses of the origin of D1 fragments indicated that the primary cleavage site of this protein was between transmembrane helices IV and V indicative of the acceptor-side mechanism for photoinhibition. The origins of other D1 protein fragments indicate that some donor-side photoinhibition may also have occurred in vivo under the conditions employed. We have shown that the spectral and functional integrating of the isolated photosystem II reaction centre complex is resistant to proteolytic cleavage by trypsin. Use of a more non-specific protease (subtilisin), however, caused significant destabilisation of the special pair of chlorophylls constituting the primary electron donor, P680, with a consequential loss of functional activity. Thus, it is possible that specific cleavage of photosystem-II reaction-centre proteins may occur in vivo following photoinhibitory damage without a significant change in structural integrity, a conclusion supported by the finding that photodamaged and normal reaction centres were isolated together.
authors.affiliation: Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, England.
authors.forename: C A
authors.initials: CA
authors.lastname: Shipton
authors.forename: J
authors.initials: J
authors.lastname: Barber
mesh_terms: D007887:Fabaceae; D008027:Light; D008970:Molecular Weight; D010449:Peptide Mapping; D010777:Photochemistry; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045332:Photosystem II Protein Complex; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D010946:Plants, Medicinal; D013057:Spectrum Analysis; D013381:Subtilisins / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D013381:Subtilisins; D014357:Trypsin