title: The glutamine synthetase from the hyperthermoacidophilic crenarcheon Sulfolobus acidocaldarius: isolation, characterization and sequencing of the gene.
abstract: The glutamine synthetase (EC 6.3.1.2) from the hyperthermoacidophilic crenarcheon Sulfolobus acidocaldarius (DSM 639) was purified to homogeneity, characterized and the glnA gene isolated and sequenced. The amount of enzyme present in the cytosolic fraction from Sulfolobus cells showed a strong variation depending on the carbon and nitrogen sources in the growth medium. The enzyme was found to be a dodecameric protein composed of identical subunits of 52 kDa. It was stable at 78 degrees C in the presence of Mn2+ ions. The catalytic activity was regulated solely by feed-back inhibition through L-alanine and glycine and not by adenylylation. No evidence for the presence of isoenzymes was found. Sequence comparison showed that the Sulfolobus protein is most closely related to the glutamine synthetases of the I-beta type despite its regulatory properties and the finding that the known euryarcheal glutamine synthetase sequences belong to the I-alpha subgroup of these enzymes. Our phylogenetic analysis suggests that the gene duplication leading to the development of the I-alpha and I-beta enzymes preceded the separation of the archea and the bacteria.
authors.affiliation: Institut für Biochemie, Medizinische Universität zu Lübeck, Germany.
authors.forename: Z
authors.initials: Z
authors.lastname: Yin
authors.forename: W G
authors.initials: WG
authors.lastname: Purschke
authors.forename: G
authors.initials: G
authors.lastname: Schäfer
authors.forename: C L
authors.initials: CL
authors.lastname: Schmidt
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002384:Catalysis; D004269:DNA, Bacterial; D004795:Enzyme Stability; D005974:Glutamate-Ammonia Ligase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D016945:Sulfolobus acidocaldarius / Q000201:enzymology*
chemical_list: D004269:DNA, Bacterial; D005974:Glutamate-Ammonia Ligase