Matsuura Shun-ichi, Itoh Tetsuji, Ishii Ryo, Sakaguchi Kengo, Tsunoda Tatsuo, Hanaoka Takaaki, Mizukami Fujio
Research Center for Compact Chemical Process, National Institute of Advanced Industrial Science and Technology, Nigatake 4-2-1, Miyagino-ku, Sendai 983-8551, Japan.
Bioconjug Chem. 2008 Jan;19(1):10-4. doi: 10.1021/bc700340e. Epub 2007 Nov 30.
The assembly and proper alignment of two heterofluorescent proteins (sGFP and DsRed) in the mesoporous channels of ethanol-treated FSM6.2 (a folded-sheet mesoporous material with a pore diameter of 6.2 nm) was confirmed using a fluorescence resonance energy transfer (FRET) technique. The sGFP-DsRed-FSM6.2 conjugate showed a large decrease in the emission of donor (sGFP) fluorescence, indicating that the conjugate functions as an energy transfer system through the combination of the two heteroproteins, due to the successful encapsulation of the sGFP-DsRed pairs in the mesopores. Fluorescence spectral analysis demonstrated that the proteins were highly dispersed and homogeneously encapsulated in the mesopores of FSM6.2, even at high concentration, although they spontaneously aggregated and showed a red shift in solution at the concentration corresponding to that in the conjugate. Furthermore, an increase in the amount of sGFP and DsRed adsorbed to the pores of FSM6.2 led to a decrease in the distance between these proteins, resulting in enhancement of FRET efficiency.
利用荧光共振能量转移（FRET）技术证实了两种异源荧光蛋白（绿色荧光蛋白和红色荧光蛋白）在乙醇处理的FSM6.2（一种孔径为6.2nm的折叠片状介孔材料）的介孔通道中组装并正确排列。绿色荧光蛋白-红色荧光蛋白-FSM6.2共轭物显示供体（绿色荧光蛋白）荧光发射大幅下降，这表明该共轭物通过两种异源蛋白的结合起到能量转移系统的作用，这是由于绿色荧光蛋白-红色荧光蛋白对成功封装在介孔中。荧光光谱分析表明，即使在高浓度下，蛋白质也高度分散且均匀地封装在FSM6.2的介孔中，尽管它们在溶液中会自发聚集，并且在与共轭物中浓度相当的浓度下会出现红移。此外，吸附到FSM6.2孔中的绿色荧光蛋白和红色荧光蛋白数量增加导致这些蛋白质之间的距离减小，从而提高了FRET效率。
