Chloroplast phosphoproteins: phosphorylation of a 12-kDa stromal protein by the redox-controlled kinase of thylakoid membranes.
Activation of the redox-controlled protein kinase of thylakoid membranes is detectable in vivo by measuring radioisotope incorporation into the light-harvesting Chl a/b protein and four photosystem II proteins (8.3, 32, 34, and 44 kDa). In normal barley leaves, the kinase is active under both aerobic and anaerobic (N2) conditions, but in the Chl b-less chlorina f2 mutant it is active only under anaerobic conditions. The responsiveness of this enzyme in the mutant to changes in the gas phase has been exploited to distinguish its protein substrates from those of other leaf protein kinases. Most of the soluble phosphoproteins of normal and mutant leaves (including a conspicuously labeled 67-kDa polypeptide) are labeled equally under both aerobic and anaerobic conditions, indicating that they are not substrates of the redox-controlled protein kinase. The major exception is a 12-kDa phosphoprotein, which is labeled in the mutant only under anaerobic conditions. The 67- and 12-kDa phosphoproteins are located in the chloroplast and are labeled when isolated organelles are incubated with [32P]orthophosphate in the light. When thylakoids and stroma are prepared from chloroplasts and are incubated with [gamma-32P]ATP in vitro, the 12-kDa protein is phosphorylated in the thylakoid preparation and then released from the membranes into the medium. The electron transport inhibitor diuron blocks activation of the redox-controlled kinase and prevents phosphorylation of the 12-kDa protein, which is thus the first example of a soluble protein to be phosphorylated by the thylakoid-bound protein kinase. The 67-kDa protein is phosphorylated by a distinct stromal kinase whose activity is not sensitive to diuron.
D000332:Aerobiosis; D000596:Amino Acids / Q000378:metabolism; D000693:Anaerobiosis; D002736:Chloroplasts / Q000201:enzymology*; D004237:Diuron / Q000494:pharmacology; D004579:Electron Transport / Q000187:drug effects; D001467:Hordeum / Q000235:genetics; D007425:Intracellular Membranes / Q000201:enzymology; D008970:Molecular Weight; D009154:Mutation; D010084:Oxidation-Reduction; D010710:Phosphates / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D010940:Plant Proteins / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
D000596:Amino Acids; D010710:Phosphates; D010750:Phosphoproteins; D010940:Plant Proteins; D004237:Diuron; D011494:Protein Kinases