Cross-linking with diimidates of glutamine synthetase from Bacillus stearothermophilus.
Glutamine synthetase [EC 6.3.2.1] from Bacillus stearothermophilus was modified with diethyl malonimidate (DEM), dimethyl adipimidate (DMA), and dimethyl suberimidate (DMS). DMA modified most epsilon-amino groups. On modification with DMA, formation of 3 to 4 cross-links/subunit resulted in a large increase in thermostability. The activity, allosteric properties and fluorescence spectrum of the enzyme were not changed on cross-linking. The SDS-polyacrylamide gel electrophoretic profiles of DEM-, DMA-, and DMS-modified enzymes suggested that the interaction berween six subunits in each of the two hexagonal rings of the protein are heterologous and are different from those between the piled subunits on different rings.
D000494:Allosteric Regulation; D000495:Allosteric Site; D003432:Cross-Linking Reagents; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D007094:Imides; D046911:Macromolecular Substances; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D003432:Cross-Linking Reagents; D007094:Imides; D046911:Macromolecular Substances; D005974:Glutamate-Ammonia Ligase