Glutamine synthetase from Mycobacterium avium.
Mycobacterium avium was previously shown to be dependent upon ammonia or glutamine as a nitrogen source. In an effort to assess the physiology of ammonia assimilation by M. avium, a characterization of its glutamine synthetase was performed. The enzyme from M. avium was purified by streptomycin sulfate treatment, ammonium sulfate precipitation, and affinity chromatography. The enzyme was unusual in that it had a pH optimum of 6.4 and maximum enzyme activity was obtained between 50 and 60 degrees C as shown by the transferase assay. The glutamine synthetase activity from batch-cultured cells decreased with increasing concentration of ammonium chloride in the range of 0.25-5 mumol/mL of medium, which demonstrated a response to environmental supply of a nitrogen source. The mycobacterial enzyme was similar to the other bacterial glutamine synthetases in terms of molecular weight and sedimentation coefficient which were 600 000 and 19.5 S, respectively, and enzyme activity was lost by treatment with a glutamate analog, methionine sulfoximine. The isoelectric point was, however, pH 4.5. Treatment of the enzyme with snake venom phosphodiesterase resulted in an increase in specific activity. AMP was released by the phosphodiesterase treatment, thus demonstrating that M. avium glutamine synthetase was regulated by adenylylation modification.
D000643:Ammonium Chloride / Q000494:pharmacology; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005974:Glutamate-Ammonia Ligase / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D005973:Glutamine / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008717:Methionine Sulfoximine / Q000494:pharmacology; D008970:Molecular Weight; D009161:Mycobacterium / Q000201:enzymology*; D009162:Mycobacterium avium / Q000201:enzymology*; D010455:Peptides / Q000032:analysis; D013696:Temperature
D005971:Glutamates; D010455:Peptides; D000643:Ammonium Chloride; D005973:Glutamine; D008717:Methionine Sulfoximine; D018698:Glutamic Acid; D005974:Glutamate-Ammonia Ligase