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1.A.5 (a) YcgF

  The YcgF protein from Escherichia coli is composed of an N-terminal BLUF domain and a C-terminal EAL domain, which contains an abundance of the amino acids, glutamine (E), alanine (A), and leucine (L). The EAL domain has been shown to encode the enzymatic activity involved in the hydrolysis of cyclic diguanosine monophosphate (c-di-GMP), which is a regulatory signaling molecule for various biological events. Therefore, it has been proposed that YcgF functions as a blue-light regulated phosphodiesterase (Blrp). The photochemical reaction dynamics of YcgF, a BLUF protein, were investigated by the pulsed laser-induced transient grating (TG) technique. The TG signal showed three reaction time constants: 2.7 micros, 13 micros, and 2 ms. The fastest was tentatively attributed to relaxation of the excited triplet state of the chromophore, flavin adenine dinucleotide (FAD), and the others represented conformational changes of the protein. The TG signal provided clear evidence that the diffusion coefficient (D) of the photo-product (3.8  10-11 m2s-1) was significantly less than that of the reactant (8.3  10-11 m2s-1), with a time constant of 2 ms at a protein concentration of 700 microM. Interestingly, the rate constant increased in proportion to the concentration of the protein, indicating that protein dimerization was one of the main reactions occurring after photoexcitation. The significant reduction in D indicates that a conformational change leading to an increase in interactions with water molecules occurs upon formation of the signaling state. The 13 microsec dynamics was attributed to the conformational change (diffusion sensitive conformational change) that induced transient dimerization. This conformational change might be an essential process for the creation of the signaling state. A detailed scheme for the photochemical reaction of YcgF is proposed.


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photo-physical-chemistry lab,京都大学大学院理学研究科 化学専攻 光物理化学研究室

〒606-8502
Kitashirakawaoiwakecho
Sakyoku, Kyoto, Japan
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