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首页> 外文期刊>Biochemistry >Femtosecond and Picosecond Dynamics of Recombinant Bacteriorhodopsin Primary Reactions Compared to the Native Protein in Trimeric and Monomeric Forms
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Femtosecond and Picosecond Dynamics of Recombinant Bacteriorhodopsin Primary Reactions Compared to the Native Protein in Trimeric and Monomeric Forms

机译：与三聚体和单体形式的天然蛋白相比，重组细菌的Femtosecond和PicoSecond动态的初级反应

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摘要

Photochemical reaction dynamics of the primary events in recombinant bacteriorhodopsin (bR(rec)) was studied by femtosecond laser absorption spectroscopy with 25-fs time resolution. bR(rec) was produced in an Escherichia coli expression system. Since bR(rec) was prepared in a DMPC-CHAPS micelle system in the monomeric form, its comparison with trimeric and monomeric forms of the native bacteriorhodopsin (bR(trim) and bR(mon), respectively) was carried out. We found that bR(rec) intermediate I (excited state of bR) was formed in the range of 100 fs, as in the case of bR(trim) and bR(mon). Further processes, namely the decay of the excited state I and the formation of intermediates J and K of bR(rec), occurred more slowly compared to bR(trim), but similarly to bR(mon). The lifetime of intermediate I, judging from the signal of Delta A(ESA) (470-480 nm), was 0.68 ps (78%) and 4.4 ps (22%) for bR(rec), 0.52 ps (73%) and 1.7 ps (27%) for bR(mon), and 0.45 ps (90%) and 1.75 ps (10%) for bR(trim). The formation time of intermediate K, judging from the signal of Delta A(GSA)(625-635 nm), was 13.5 ps for bR(rec), 9.8 ps for bR(mon), and 4.3 ps for bR(trim). In addition, there was a decrease in the photoreaction efficiency of bR(rec) and bR(mon) as seen by a decrease in absorbance in the differential spectrum of the intermediate K by similar to 14%. Since photochemical properties of bR(rec) are similar to those of the monomeric form of the native protein, bR(rec) and its mutants can be considered as a basis for further studies of the mechanism of bacteriorhodopsin functioning.
机译：通过Femtosecond激光吸收光谱研究了重组烧伤素（BR（REC））中的主要事件的光化学反应动力学，具有25-FS时间分辨率。 BR（REC）是在大肠杆菌表达系统中产生的。由于在单体形式中在DMPC-CHAPS胶束系统中制备BR（REC），因此进行其与三聚体和单体形式的天然Bacteriorhodopsin（Br（Trim）和Br（Mon）的单体形式的比较。我们发现BR（REC）中间I（BR的激发状态）在100 fs的范围内形成，如BR（修剪）和BR（MON）的情况。进一步的方法，即激发态I的衰减和Br（Rec）的中间体J和K的形成，与Br（修剪）相比，更缓慢地发生，但与Br（mon）类似。从Δα（ESA）的信号（470-480nm）判断中间I的寿命为BR（REC）的0.68 ps（78％）和4.4 ps（22％），0.52 ps（73％）和Br（MON）的1.7 ps（27％），Br（mon）和0.45 ps（90％）和1.75 ps（10％）的br（修剪）。中间k的形成时间，从Delta A（GSA）（625-65nm）的信号判断为13.5 ps，对于Br（Mon）的Br（Rec），9.8 ps，Br（修剪）的4.3 ps。另外，Br（Rec）和Br（mon）的光反应效率降低，通过中间k的差分谱的吸光度降低，与14％相似。由于Br（Rec）的光化学性质类似于天然蛋白质的单体形式的化学性质，因此Br（Rec）及其突变体可以被认为是进一步研究Bacteriorhodopsin功能的机制的基础。

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