Maintainance of specificity, information, and thermostability in thermophilic Bacillus sp. glutamine synthetase.
Glutamine synthetase has been purified to homogeneity from B. subtilis (37 degrees) B. stearothermophilus (55 degrees), and B. caldolyticus (75 degrees). Those characteristics compared include size (6.0 +/- 0.3 X 10(5) daltons), quaternary structure (12 SU) amino acid content, substrate Km's and specificity for structural analogs, metal ion activation, number and kind of separate feedback modifier sites, and the complexity of modifier-substrate and modifier-modifier site interactions. Although the 37 degrees and 55 degrees systems are quite similar, the 75 degrees system shows important alterations in substrate specificity and modes of modifier action. Whereas at 37 degrees and 55 degrees AMP inhibits synergistically with amino acids (glycine, glutamine, histidine), the 75 degrees enzyme is inhibited directly by the products ADP, (which assumes the role of AMP) and glutamine, plus other ligands. Ligand binding domains are compared and found to be very different. Thermostabilization occurs by (a) protection by bound L-glutamate, (b) protein aggregation, (c) trends in the content of total polar residues, total Asx + Flx residues, the average hydrophobicity, and (d) disulfide bond cross-linking. Such studies provide insights to molecular evolution occurring with changes in environmental stress.
D000227:Adenine Nucleotides / Q000494:pharmacology; D000596:Amino Acids / Q000032:analysis; D001407:Bacillus / Q000201:enzymology*; D001412:Bacillus subtilis / Q000201:enzymology; D002413:Cations, Divalent / Q000494:pharmacology; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D005974:Glutamate-Ammonia Ligase / Q000032:analysis / Q000378:metabolism; D005973:Glutamine / Q000494:pharmacology; D005998:Glycine / Q000494:pharmacology; D006639:Histidine / Q000494:pharmacology; D006358:Hot Temperature; D007700:Kinetics; D008024:Ligands; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D011487:Protein Conformation
D000227:Adenine Nucleotides; D000596:Amino Acids; D002413:Cations, Divalent; D008024:Ligands; D005973:Glutamine; D008345:Manganese; D006639:Histidine; D005974:Glutamate-Ammonia Ligase; D008274:Magnesium; D005998:Glycine