Long-Range Energy Transfer in Protein Megamolecules.
Long-Range Energy Transfer in Protein Megamolecules.
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DOI：
10.1021/jacs.8b08208
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发表时间：
2018-11-21
影响因子：
15
通讯作者：
Goodson T 3rd
中科院分区：
文献类型：
--
作者：
Taylor EL;Metcalf KJ;Carlotti B;Lai CT;Modica JA;Schatz GC;Mrksich M;Goodson T 3rd
在这项调查中，我们报告的证据，在新的蛋白质为基础的巨分子与供体和受体荧光蛋白之间的距离可调的能量转移。本研究中使用的巨分子是单分散性低聚物，分子量约为100-300 kDa，长度约为5-20 nm，是融合蛋白构建块和共价交联剂的精确定义结构。这样的结构是有前途的，因为蛋白质复合物中的能量转移的研究通常是困难的，在这个长的长度制度，由于合成的限制。我们将荧光蛋白掺入巨分子结构中，并通过改变二聚体缀合物中交联剂的长度和插入非荧光间隔蛋白来改变供体和受体之间的间隔距离以产生寡聚体。双光子吸收测量表明在巨分子中供体和受体偶极之间存在强耦合。对于二聚体系统，没有影响的交联剂的长度上的能量转移效率观察到与稳态荧光调查。然而，对于相同的二聚体缀合物，能量转移速率降低后，增加交联剂的长度，通过荧光上转换评价。分子动力学模拟用于合理化结果，提供稳态结果的测量和计算能量转移长度之间的定量一致性，并表明时间分辨测量和稳态测量之间的差异源于大尺度波动的长时间尺度巨分子结构。我们的研究结果表明，随着巨分子尺寸的增加，能量传递长度增加。这是大蛋白质巨分子中长距离能量转移的证据。
In this investigation, we report evidence for energy transfer in new protein-based megamolecules with tunable distances between donor and acceptor fluorescent proteins. The megamolecules used in this work are monodisperse oligomers, with molecular weights of ~100–300 kDa and lengths of ~5–20 nm, and are precisely defined structures of fusion protein building blocks and covalent cross-linkers. Such structures are promising because the study of energy transfer in protein complexes is usually difficult in this long length regime due to synthetic limitations. We incorporated fluorescent proteins into the megamolecule structure and varied the separation distance between donor and acceptor by changing the length of the cross-linker in dimer conjugates and inserting nonfluorescent spacer proteins to create oligomers. Two-photon absorption measurements demonstrated strong coupling between donor and acceptor dipoles in the megamolecules. For the dimer systems, no effect of the crosslinker length on energy transfer effciency was observed with the steady-state fluorescence investigation. However, for the same dimer conjugates, energy transfer rates decreased upon increasing cross-linker length, as evaluated by fluorescence up-conversion. Molecular dynamics simulations were used to rationalize the results, providing quantitative agreement between measured and calculated energy transfer lengths for steady-state results, and showing that the differences between the time-resolved and steady-state measurements arise from the long time scale for large-scale fluctuations in the megamolecule structure. Our results show an increase in energy transfer length with increasing megamolecule size. This is evidence for long-range energy transfer in large protein megamolecules.
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影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ
DOI：
10.1021/acs.jpcc.6b11235
发表时间：
2017-01-26
期刊：
The journal of physical chemistry. C, Nanomaterials and interfaces
影响因子：
--
作者：
Blacker TS;Chen W;Avezov E;Marsh RJ;Duchen MR;Kaminski CF;Bain AJ
通讯作者：
Bain AJ
影响因子：
5.5
作者：
Goetz, Andreas W.;Williamson, Mark J.;Xu, Dong;Poole, Duncan;Le Grand, Scott;Walker, Ross C.
通讯作者：
Walker, Ross C.
影响因子：
3.3
作者：
Cunningham, Paul D.;Khachatrian, Ani;Melinger, Joseph S.
通讯作者：
Melinger, Joseph S.
影响因子：
4
作者：
Doan, Phi;Pitter, Demar R. G.;Goodson, Theodore, III
通讯作者：
Goodson, Theodore, III