Huppert Dan, Leiderman Pavel, Ben-Ziv Moran, Genosar Liat, Cohen Lior
Raymond and Beverly Sackler Faculty of Exact Sciences, School of Chemistry, Tel Aviv University, Tel Aviv 69978, Israel.
J Phys Chem B. 2005 Mar 10;109(9):4241-51. doi: 10.1021/jp0443153.
Picosecond time-correlated single-photon counting was used to measure the proton-transfer rate of green fluorescent protein (GFP) excited by several wavelengths between 266 and 405 nm. When samples of GFP in water and D2O are excited at short wavelengths, lambda(ex) < 295 nm, the fluorescence properties are largely modified with respect to excitation at a wavelength around 400 nm, the peak of the absorption band of the S0 --> S1 transition of the ROH form of the chromophore. The shorter the excitation wavelength, the longer the decay time of the ROH emission band at 450 nm and the longer the rise time of the RO- emission band at 512 nm. The proton transfer is slower by an order of magnitude and about a factor of 3 when GFP in water and D2O are excited by 266 nm, respectively.
皮秒时间相关单光子计数被用于测量绿色荧光蛋白（GFP）在266至405纳米之间多个波长激发下的质子转移速率。当水中和重水中的GFP样品在短波长（λex < 295纳米）激发时，相对于在约400纳米波长（发色团ROH形式的S0→S1跃迁吸收带的峰值）激发，其荧光特性有很大改变。激发波长越短，450纳米处ROH发射带的衰减时间越长，512纳米处RO-发射带的上升时间越长。当水中和重水中的GFP分别被266纳米激发时，质子转移速度慢一个数量级，且大约慢3倍。
