Journal
BIOORGANIC & MEDICINAL CHEMISTRY
Volume 17, Issue 20, Pages 7381-7386Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.bmc.2009.08.015
Keywords
Biosensor; PH domain; Structure-based design; Green fluorescent protein; Inositol polyphosphates
Funding
- Ministry of Education, Culture, Sports, Science and Technology, Japan [19021023, 20241051]
- Grants-in-Aid for Scientific Research [19021023, 20241051, 20249015] Funding Source: KAKEN
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A fluorescent sensor for the detection of inositol-1,3,4,5-tetrakisphosphate, Ins(1,3,4,5)P-4, was constructed from a split PH domain and a single circularly permuted GFP. A structure-based design was conducted to transduce a ligand-induced subtle structural perturbation of the split PH domain to an alteration in the population of the protonated and the deprotonated states of the GFP chromophore. Excitation of each distinct absorption band corresponding to the protonated or the deprotonated state of GFP resulted an increase and a decrease, respectively, in the intensity of emission spectra upon addition of Ins(1,3,4,5)P-4 to the split PH domain-based sensor. The Ins(1,3,4,5)P-4 sensor retained the ligand affinity and the selectivity of the parent PH domain, and realized the ratiometric fluorescence detection of Ins(1,3,4,5)P-4. (c) 2009 Elsevier Ltd. All rights reserved.
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