title: Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of surface residue Val433.
abstract: After thermal incubation at 48 °C for 10 min, single variants of Moloney murine leukemia virus reverse transcriptase, V433R and V433K in which a surface hydrophobic residue, Val433, was mutated, retained 55% of initial reverse transcription activity, while the wild-type enzyme retained 17%. After thermal incubation at 50 °C for 10 min, multiple variants D108R/E286R/V433R and D108R/E286R/V433R/D524A, in which Val433→Arg was combined with stabilizing mutations we identified previously, Asp108→Arg and Glu286→Arg, and RNase H activity-eliminating mutation Asp524→Ala, retained 70% of initial activity, exhibiting higher stability than V433R or V433K.
authors.affiliation: a Division of Food Science and Biotechnology, Graduate School of Agriculture , Kyoto University , Kyoto , Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Konishi
authors.forename: Xiaochen
authors.initials: X
authors.lastname: Ma
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Yasukawa
mesh_terms: D004795:Enzyme Stability / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D008979:Moloney murine leukemia virus / Q000201:enzymology*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011487:Protein Conformation; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry* / Q000235:genetics*; D013696:Temperature; D014633:Valine
chemical_list: D012194:RNA-Directed DNA Polymerase; D014633:Valine
keywords: Moloney murine leukemia virus; reverse transcriptase; site-directed mutagenesis; surface hydrophobic residue; thermostability