Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of the surface residue Val433
Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of the surface residue Val433
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通过表面残基 Val433 的定点诱变稳定莫洛尼鼠白血病病毒逆转录酶
DOI：
10.1080/09168451.2014.877186
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
保川 清
中科院分区：
文献类型：
--
作者：
小西 篤;馬 暁晨;保川 清
在48 °C下热孵育10分钟后，其中表面疏水残基Val 433突变的莫洛尼鼠白血病病毒逆转录酶的单一变体V433 R和V433 K保留了55%的初始逆转录活性，而野生型酶保留了17%。在50 °C下热孵育10分钟后，多个变体D108 R/E286 R/V433 R和D108 R/E286 R/V433 R/D524 A，其中Val 433 →Arg与我们先前鉴定的稳定突变Asp 108 →Arg和Glu 286 →Arg以及RNase H活性消除突变Asp 524 →Ala组合，保留了70%的初始活性，表现出比V433 R或V433 K更高的稳定性。
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.