title: Increased functional activity of elongation factor G with G16V mutation in the GTP-binding domain.
abstract: Oligonucleotide-directed mutagenesis was used to obtain elongation factor G from Thermus thermophilus with the G16V mutation in its GTP-binding domain. Functional studies of the mutated protein and elongation factor G from E. coli were carried out. The data revealed that the G16V mutant retains high thermostability, has an increased ribosome-dependent GTPase activity, and its translation activity in cell-free translation system is equal to that of the factor G from E. coli. The mutated protein with an uncleavable GTP analog also has an increased affinity to the ribosomes.
authors.affiliation: Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia.
authors.forename: K A
authors.initials: KA
authors.lastname: Martemyanov
authors.forename: A
authors.initials: A
authors.lastname: Liljas
authors.forename: A T
authors.initials: AT
authors.lastname: Gudkov
mesh_terms: D001483:Base Sequence; D020134:Catalytic Domain / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D020963:GTP Phosphohydrolase-Linked Elongation Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005798:Genes, Bacterial; D006160:Guanosine Triphosphate / Q000378:metabolism*; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D016963:Thermus thermophilus / Q000235:genetics / Q000378:metabolism
chemical_list: D017931:DNA Primers; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; D011994:Recombinant Proteins; D006160:Guanosine Triphosphate; D020963:GTP Phosphohydrolase-Linked Elongation Factors