Creemers T M, Lock A J, Subramaniam V, Jovin T M, Völker S
Center for the Study of the Excited States of Molecules, Huygens and Gorlaeus Laboratories, University of Leiden, The Netherlands.
Nat Struct Biol. 1999 Jun;6(6):557-60. doi: 10.1038/9335.
Several studies have led to the conclusion that, in the green fluorescent protein (GFP) of the jellyfish Aequorea victoria, a photoconversion involving excited-state proton transfer occurs from an A- to a B-form, while an intermediate I-form was held responsible for the green fluorescence. Here we have identified the I-form of wild-type GFP in absorption, located the 0-0 transitions of all three forms A, B and I, and determined vibrational frequencies of the ground and excited states. The intrinsically narrow 0-0 transitions are revealed by the wavelengths at which holes can be burnt. The pathways of photointerconversion are unraveled by excitation, emission and hole-burning spectroscopy. We present an energy-level scheme that has significant implications for GFP-mutants, which likewise can occur in the three photo-interconvertible forms.
多项研究得出结论，在维多利亚多管水母的绿色荧光蛋白（GFP）中，涉及激发态质子转移的光转换从A形式发生到B形式，而中间的I形式则产生绿色荧光。在此，我们通过吸收鉴定了野生型GFP的I形式，确定了A、B和I三种形式的0-0跃迁，并测定了基态和激发态的振动频率。本质上狭窄的0-0跃迁通过可烧孔的波长得以揭示。光互变途径通过激发、发射和烧孔光谱得以阐明。我们提出了一个能级方案，该方案对同样可以以三种光互变形式出现的GFP突变体具有重要意义。
