Purification and regulation of glutamine synthetase in a collagenolytic Vibrio alginolyticus strain.
Glutamine synthetase (EC 6.3.1.2) has been purified from a collagenolytic Vibrio alginolyticus strain. The apparent molecular weight of the glutamine synthetase subunit was approximately 62,000. This indicates a particle weight for the undissociated enzyme of 744,000, assuming the enzyme is the typical dodecamer. The glutamine synthetase enzyme had a sedimentation coefficient of 25.9 S and seems to be regulated by adenylylation and deadenylylation. The pH profiles assayed by the gamma-glutamyltransferase method were similar for NH4-shocked and unshocked cell extracts and isoactivity point was not obtained from these curves. The optimum pH for purified and crude cell extracts was 7.9. Cell-free glutamine synthetase was inhibited by some amino acids and AMP. The transferase activity of glutamine synthetase from mid-exponential phase cells varied greatly depending on the sources of nitrogen or carbon in the growth medium. Glutamine synthetase level was regulated by nitrogen catabolite repression by (NH4)2SO4 and glutamine, but cells grown in the presence of proline, leucine, isoleucine, tryptophan, histidine, glutamic acid, glycine and arginine had enhanced levels of transferase activity. Glutamine synthetase was not subject to glucose, sucrose, fructose, glycerol or maltose catabolite repression and these sugars had the opposite effect and markedly enhanced glutamine synthetase activity.
Bodasing
S J
SJ
Brandt
P W
PW
Robb
F T
FT
Woods
D R
DR
D000645:Ammonium Sulfate / Q000494:pharmacology; D002244:Carbon / Q000494:pharmacology; D005246:Feedback; D005974:Glutamate-Ammonia Ligase / Q000032:analysis / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D009584:Nitrogen / Q000494:pharmacology; D014733:Vibrio / Q000201:enzymology*
D002244:Carbon; D005974:Glutamate-Ammonia Ligase; D008274:Magnesium; D009584:Nitrogen; D000645:Ammonium Sulfate