Direct observation of motor protein stepping in living cells using MINFLUX.

Direct observation of motor protein stepping in living cells using MINFLUX.
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DOI:
10.1126/science.ade2676
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发表时间:
2023-03-10
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Ries J
Ries J
中科院分区:
其他
文献类型:
--
作者:
Deguchi T;Iwanski MK;Schentarra EM;Heidebrecht C;Schmidt L;Heck J;Weihs T;Schnorrenberg S;Hoess P;Liu S;Chevyreva V;Noh KM;Kapitein LC;Ries J

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分子机器的动态测量可以提供对其机制的宝贵见解,但在活细胞中一直具有挑战性。在这里,我们利用最近推出的超分辨率技术MINFLUX开发了二维和三维纳米空间和毫秒时间分辨率的单个荧光团的活细胞跟踪。利用这种方法,我们解决了运动蛋白激酶1在活细胞微管上行走时的精确步进运动。在固定细胞微管上行走的马达的纳米级跟踪也使我们能够用原丝分辨率解析微管细胞骨架的结构。
Dynamic measurements of molecular machines can provide invaluable insights into their mechanism but have been challenging in living cells. Here, we developed live-cell tracking of single fluorophores with nanometer spatial and millisecond temporal resolution in two and three dimensions using the recently introduced super-resolution technique MINFLUX. Using this approach, we resolved the precise stepping motion of the motor protein Kinesin-1 as it walks on microtubules in living cells. Nanoscopic tracking of motors walking on the microtubules of fixed cells also enabled us to resolve the architecture of the microtubule cytoskeleton with protofilament resolution.
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