title: Phosphorylation and Dephosphorylation of Guard-Cell Proteins from Vicia faba L. in Response to Light and Dark.
abstract: Phosphorylation and dephosphorylation of proteins were investigated in guard-cell protoplasts from Vicia faba L. When guard-cell protoplasts were incubated with 32Pi in the dark for 80 min, several proteins, with molecular masses of 42, 40, 34, 32, 26, and 19 kD, were phosphorylated. Illumination of the dark-adapted protoplasts with red light caused dephosphorylation of the 26-kD protein, but there was no detectable change in levels of phosphorylation in other proteins. In the dephosphorylation of the 26-kD protein, far-red light of 730 nm was most effective, but when the light was turned off, the protein was phosphorylated to the original level within 10 min. Subcellular fractionation of guard-cell protoplasts indicated that the 26-kD protein was located in the chloroplast. The migration pattern of the 26-kD protein was exactly the same as the light-harvesting Chl a/b protein complex of photosystem II (LHCPII) from Vicia mesophyll cells on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The dephosphorylated 26-kD protein was phosphorylated by adding sodium hydrosulfite, a strong reducing agent, under the far-red illumination of guard-cell protoplasts. The magnitude of dephosphorylation by red light (660 nm) was increased by 3-(3,4-dichlorophenyl)-1,1-dimethylurea, an electron transfer inhibitor of photosystem II (PSII). Light-induced dephosphorylation was inhibited by 1 nM okadaic acid, an inhibitor of serine/threonine protein phosphatase. From these results, it is concluded that the 26-kD protein is LHCPII and that LHCPII is present mostly in the phosphorylated form in the dark and is dephosphorylated by type 2A protein phosphatase under the light absorbed by photosystem I in Vicia guard-cell protoplasts.
authors.affiliation: Department of Biology, Faculty of Science, Kyushu University, Hakozaki, Fukuoka, 812 Japan (T.K., M.N.).
authors.forename: T.
authors.initials: T
authors.lastname: Kinoshita
authors.forename: Ki.
authors.initials: K
authors.lastname: Shimazaki
authors.forename: M.
authors.initials: M
authors.lastname: Nishimura