Isolation and crystallization of unadenylylated glutamine synthetase from Salmonella typhimurium.
The enzyme glutamine synthetase (GS) has been isolated from a mutant strain of Salmonella typhimurium, constructed by Kustu, which lacks the enzymatic activity for adenylylation of glutamine synthetase. Thus the purified GS is uniformly unadenylylated, as confirmed by gel electrophoresis and enzyme assays. It crystallizes readily in many morphologies, at least six of which are distinct polymorphs. The most favorable crystal form for structural studies belongs to space group C2, with unit cell dimensions a = 235.5 A, b = 134.5 A, c = 200.1 A, beta = 102.8 degrees, and with one GS molecule per asymmetric unit. The crystals diffract to about 2.8 A resolution in rotation X-ray photographs and thus appear suitable for structural studies at moderate resolution. These crystals are isomorphous with crystalline GS from Escherichia coli in both adenylylated and unadenylylated states, suggesting that the enzymes from the two bacteria are similar molecules, and that adenylylation does not greatly affect the conformation of the molecule.
D000249:Adenosine Monophosphate; D055598:Chemical Phenomena; D002621:Chemistry; D003460:Crystallization; D005974:Glutamate-Ammonia Ligase / Q000302:isolation & purification*; D011487:Protein Conformation; D012486:Salmonella typhimurium / Q000201:enzymology*; D014961:X-Ray Diffraction
D000249:Adenosine Monophosphate; D005974:Glutamate-Ammonia Ligase