Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of surface residue Val433

作者：Konishi Atsushi; Ma Xiaochen; Yasukawa Kiyoshi^*

来源：Bioscience Biotechnology and Biochemistry, 2014, 78(1): 75-78.

DOI：10.1080/09168451.2014.877186

• thermostability
• surface hydrophobic residue
• site-directed mutagenesis
• reverse transcriptase
• Moloney murine leukemia virus

After thermal incubation at 48 degrees 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 degrees 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.

• 出版日期2014-1-2