阿姆斯特丹自由大學(xué)(VU University Amsterdam)的 Erwin J.G. Peterman在Chemical Society Reviews上發(fā)表了熒光共振能量轉(zhuǎn)移和驅(qū)動(dòng)蛋白分子的綜述。
Abstract:Förster Resonance Energy Transfer (FRET) is the phenomenon of non-radiative transfer of electronic excitations from a donor fluorophore to an acceptor, mediated by electronic dipole–dipole coupling. The transfer rate and, as a consequence, efficiency depend non-linearly on the distance between the donor and the acceptor. FRET efficiency can thus be used as an effective and accurate reporter of distance between two fluorophores and changes thereof. Over the last 50 years or so, FRET has been used as a spectroscopic ruler to measure conformations and conformational changes of biomolecules. More recently, FRET has been combined with microscopy, ultimately allowing measurement of FRET between a single donor and a single acceptor pair. In this review, we will explain the physical foundations of FRET and how FRET can be applied to biomolecules. We will highlight the power of the different FRET approaches by focusing on its application to the motor protein kinesin, which undergoes several conformational changes driven by enzymatic action, that ultimately result in unidirectional motion along microtubule filaments, driving active transport in the cell. Single-molecule and ensemble FRET studies of different aspects of kinesin have provided numerous insights into the complex chemomechanical mechanism of this fascinating protein.
熒光共振能量轉(zhuǎn)移(FRET)是熒光供體到受體直接電子激發(fā)態(tài)的非輻射符合轉(zhuǎn)移,通過(guò)電子偶極-偶極聯(lián)接調(diào)節(jié)。因此,有效的轉(zhuǎn)移速率取決于供體和受體直接的非線性距離。FRET的效率可以用作兩個(gè)熒光基團(tuán)距離和變化實(shí)際和精確的監(jiān)控器。在過(guò)去的50年里,F(xiàn)RET已經(jīng)被用作生物分子構(gòu)造和構(gòu)型變化的“光譜標(biāo)尺”。而且最近,F(xiàn)RET和顯微鏡技術(shù)結(jié)合,最終測(cè)量單分子供體和單分子受體對(duì)直接的FRET效率。
在本文中,我們解釋了FRET的物理基礎(chǔ),以及FRET技術(shù)如何應(yīng)用于生物分子。在驅(qū)動(dòng)蛋白分子應(yīng)用方面,我們強(qiáng)調(diào)了不同F(xiàn)RET方法的差別,主要是由于酶催化活性導(dǎo)致的不同的構(gòu)象變化,最終導(dǎo)致沿著微管細(xì)絲的單向性的運(yùn)動(dòng),進(jìn)而驅(qū)動(dòng)細(xì)胞內(nèi)的主動(dòng)運(yùn)輸。不同方面的驅(qū)動(dòng)蛋白的單分子和整體FRET研究為這一精妙蛋白質(zhì)的復(fù)雜的化學(xué)機(jī)械機(jī)制提供了深刻見解。 |