title: Expression of the GTPase activating domain of the neurofibromatosis type 1 (NF1) gene in Escherichia coli and role of the conserved lysine residue.
abstract: A small catalytic domain from the neurofibromatosis type 1 gene, NF1-333, consisting of 333 amino acids between residues 1197 and 1528, including an additional N-terminal methionine, was expressed in Escherichia coli as a soluble protein. Its catalytic activity under non-saturating conditions is similar to the full-length p120-GAP but different from truncated GAP-334. Under saturating conditions its kcat and KM are lower. Lys-1422, which is totally conserved in all GAP proteins, was mutated and the properties of the mutant protein investigated. Lys-1422 seems to be essential for the stability of the proteins and not for its catalytic activity.
authors.affiliation: Max-Planck-Institut für Medizinische Forschung, Abteilung Biophysik, Heidelberg, Germany.
authors.forename: L
authors.initials: L
authors.lastname: Wiesmüller
authors.forename: A
authors.initials: A
authors.lastname: Wittinghofer
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics*; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D005798:Genes, Bacterial; D016514:Genes, Neurofibromatosis 1; D006358:Hot Temperature; D007700:Kinetics; D008239:Lysine / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D015689:Oncogene Protein p21(ras) / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D020558:GTP Phosphohydrolases; D015689:Oncogene Protein p21(ras); D008239:Lysine