Impact of fluorescent protein fusions on the bacterial flagellar motor.
Impact of fluorescent protein fusions on the bacterial flagellar motor.
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DOI：
10.1038/s41598-017-11241-w
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发表时间：
2017-10-03
影响因子：
4.6
通讯作者：
Pedaci F
中科院分区：
文献类型：
--
作者：
Heo M;Nord AL;Chamousset D;van Rijn E;Beaumont HJE;Pedaci F
荧光融合蛋白打开了一个直接和独特的窗口蛋白质功能。然而，它们也引入了干扰天然蛋白质功能的风险。因此，荧光融合的成功应用依赖于仔细的评估和副作用的最小化，但许多应用仍然缺乏这种洞察力。这在研究马达蛋白的内部动力学时特别相关，其中化学和机械反应坐标都可能受到影响。荧光蛋白融合到定子的细菌鞭毛马达（BFM）先前已被用来揭开马达亚基动力学。在这里，我们报告了三种荧光蛋白融合到定子上对单个电机的影响，所有这些都改变了BFM的行为。单个定子产生的扭矩减少，而它们的化学计量保持不受影响。MotB融合降低了开关频率，并在两个方向上的速度中诱导了一种新的偏置依赖的不对称性。这些影响可以通过在融合点处插入接头来减轻。这些研究结果提供了一个定量的帐户上的定子荧光融合BFM动力学及其缓解的影响-新的见解，推进使用荧光融合探测蛋白质复合物的动力学。
Fluorescent fusion proteins open a direct and unique window onto protein function. However, they also introduce the risk of perturbation of the function of the native protein. Successful applications of fluorescent fusions therefore rely on a careful assessment and minimization of the side effects, but such insight is still lacking for many applications. This is particularly relevant in the study of the internal dynamics of motor proteins, where both the chemical and mechanical reaction coordinates can be affected. Fluorescent proteins fused to the stator of the Bacterial Flagellar Motor (BFM) have previously been used to unveil the motor subunit dynamics. Here we report the effects on single motors of three fluorescent proteins fused to the stators, all of which altered BFM behavior. The torque generated by individual stators was reduced while their stoichiometry remained unaffected. MotB fusions decreased the switching frequency and induced a novel bias-dependent asymmetry in the speed in the two directions. These effects could be mitigated by inserting a linker at the fusion point. These findings provide a quantitative account of the effects of fluorescent fusions to the stator on BFM dynamics and their alleviation— new insights that advance the use of fluorescent fusions to probe the dynamics of protein complexes.
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影响因子：
16.1
作者：
Chen, Xiaoying;Zaro, Jennica L.;Shen, Wei-Chiang
通讯作者：
Shen, Wei-Chiang
DOI：
10.1093/protein/14.8.529
发表时间：
2001-08-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Arai, R;Ueda, H;Nagamune, T
通讯作者：
Nagamune, T
影响因子：
64.8
作者：
BLOCK, SM;BERG, HC
通讯作者：
BERG, HC
DOI：
10.1073/pnas.1511691113
发表时间：
2016-04-26
影响因子：
11.1
作者：
Hosu, Basarab G.;Nathan, Vedavalli S. J.;Berg, Howard C.
通讯作者：
Berg, Howard C.
EGFP的黑暗面：多泛素化。
影响因子：
3.7
作者：
Baens, Mathijs;Noels, Heidi;Broeckx, Vicky;Hagens, Sofie;Fevery, Sabine;Billiau, An D.;Vankelecom, Hugo;Marynen, Peter
通讯作者：
Marynen, Peter