Degradation of the D1 protein of photosystem II under illumination in vivo: two different pathways involving cleavage or intermolecular cross-linking.
The D1 protein of the photosystem II reaction center turns over the most rapidly of all the proteins of the thylakoid membrane under illumination in vivo. In vitro, the D1 protein sustained cleavage in a surface-exposed loop (DE loop) or cross-linking with another reaction center protein, the D2 protein or cytochrome b(559), under illumination. We found that the D1 protein was damaged in essentially the same way in vivo, although the resultant fragments and cross-linked adducts barely accumulated due to digestion by proteases. In vitro studies detected a novel stromal protease(s) that digested the adducts but not the monomeric D1 protein. These observations suggest that, in addition to cleavage, the cross-linking reactions themselves are processes involved in complete degradation of the D1 protein in vivo. Peptide mapping experiments located the cross-linking sites with the D2 protein among residues 226-244, which includes the cross-linking site with cytochrome b(559) [Barbato, R., et al. (1995) J. Biol. Chem. 270, 24032-24037], in the N-terminal part of the DE loop, while N-terminal amino acid sequencing of the fragment located the cleavage site around residue 260 in the C-terminal part of the loop. We propose a model explaining the occurrence of simultaneous cleavage and cross-linking and discuss the mechanisms of complete degradation of the D1 protein in vivo.
Mizusawa
Naoki
N
Photosynthesis Laboratory, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan.
Tomo
Tatsuya
T
Satoh
Kimiyuki
K
Miyao
Mitsue
M
D002736:Chloroplasts / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D003432:Cross-Linking Reagents; D003573:Cytochrome b Group / Q000737:chemistry / Q000378:metabolism; D003624:Darkness; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism; D005609:Free Radicals / Q000378:metabolism; D008027:Light; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D018515:Plant Leaves / Q000378:metabolism*; D018724:Spinacia oleracea / Q000378:metabolism* / Q000528:radiation effects; D017154:Stromal Cells / Q000201:enzymology; D020524:Thylakoids / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects
D003432:Cross-Linking Reagents; D003573:Cytochrome b Group; D005609:Free Radicals; D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; C019313:cytochrome b559; D010450:Endopeptidases