国家自然科学基金(19890380-4,30240042;39730160-Ⅱ)和中国科学院生物物理研究所所长基金资助项目.
This work was supported by grants from The National Natural Sciences Foundation of China (19890380-4,30240042;39730160-Ⅱ) and The Foundationn of Institute of Biophysics, The Chinese Academy of Sciences.
从基因突变的F1-ATP酶(基因突变质粒,α-C193S, γ-S107C,β亚基带有10个组氨酸标记(His-Tag),转入到菌株大肠杆菌JM103)的菌株中筛选出一高表达菌株.该菌株表达的F1-ATP酶经纯化后其水解活性明显高于文献值. 从单分子水平上进行观察,发现在水解ATP过程中,γ亚基上连接的荧光标记蛋白微丝,其旋转速度要比文献中同样条件下快约一倍.
The high activity F1-ATPase (the mutant α-C193S, γ-S107C, a ten histidine (His) tag inserted immediately downstream of the β initiation codon, α3β3γ subcomplex) of thermophilic Bacillus PS3 was purified from E.coli. JM103 Δ(uncB-uncD),in which the majority of F1-ATPase genes have been eliminated. It was found that the enzyme hydrolyzed ATP more efficiently than previous papers. During the F1-ATPase hydrolyzing ATP, the rotary rate of the fluorescent actin filament attached to γ subunit of F1-ATPase was about one times faster than that of previous papers in the similar conditions.
王兴胜,崔元波,张英豪,乐加昌,江丕栋.高活性F1-ATP酶单分子旋转初步观察[J].生物化学与生物物理进展,2003,30(2):194-198
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