In vivo modification of Azotobacter chroococcum glutamine synthetase.
A monospecific anti-(glutamine synthetase) antibody raised against glutamine synthetase of the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 immunoreacted with glutamine synthetase from the N2-fixing heterotrophic bacterium Azotobacter chroococcum. In Western-blotting experiments this antibody recognized a single protein of a molecular mass of 59 kDa corresponding to glutamine synthetase subunit. This protein was in vivo-labelled in response to addition of ammonium, both [3H]adenine and H(3)32PO4 preincubation of the cells being equally effective. Nevertheless, the amount of glutamine synthetase present in A. chroococcum was independent of the available nitrogen source. Modified, inactive glutamine synthetase was re-activated by treatment with snake-venom phosphodiesterase but not by alkaline phosphatase. L-Methionine-DL-sulphoximine, an inhibitor of glutamine synthetase, prevented the enzyme from being covalently modified. We conclude that, in A. chroococcum, glutamine synthetase is adenylylated in response to ammonium and that for the modification to take place ammonium must be metabolized.
Muñoz-Centeno
M C
MC
Instituto de Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla-Consejo Superior de Investigaciones Científicas, Spain.
Cejudo
F J
FJ
Paneque
A
A
D000225:Adenine / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000494:pharmacology; D000918:Antibody Specificity; D001395:Azotobacter / Q000201:enzymology*; D015153:Blotting, Western; D000458:Cyanobacteria / Q000201:enzymology; D004789:Enzyme Activation; D005974:Glutamate-Ammonia Ligase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D007163:Immunosorbent Techniques; D008717:Methionine Sulfoximine / Q000494:pharmacology; D008970:Molecular Weight; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000494:pharmacology; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology
D000644:Quaternary Ammonium Compounds; D008717:Methionine Sulfoximine; D000469:Alkaline Phosphatase; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom; D005974:Glutamate-Ammonia Ligase; D000225:Adenine