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A fluorescence-based method for evaluating inositol 1,4,5-trisphosphate receptor ligands: Determination of subtype selectivity and partial agonist effects

Paper ID Volume ID Publish Year Pages File Format Full-Text
23499 43444 2013 7 PDF Available
Title
A fluorescence-based method for evaluating inositol 1,4,5-trisphosphate receptor ligands: Determination of subtype selectivity and partial agonist effects
Abstract

•We developed a new method for evaluating properties of IP3R ligands using fluorescent biosensors.•This method revealed a diversity in subtype selectivity of adenophostin A and its analogs.•We also found a compound showing a partial agonistic effect on IP3R1.

Inositol 1,4,5-trisphosphate (IP3) receptors consist of three subtypes: IP3R1, IP3R2, and IP3R3. Although numerous IP3 receptor ligands have been synthesized, none of the subtype-selective ligands are known. We have developed a simple fluorescence method to examine the subtype selectivity of IP3 receptor ligands using FRET-based IP3 biosensors LIBRAvI, LIBRAvII, and LIBRAvIII. The addition of IP3 or adenophostin A (ADA) to permeabilized biosensor-expressing cells increased the fluorescence ratios of these biosensors in a concentration-dependent manner, and the potency of ADA relative to that of IP3 in terms of the changes in the fluorescence ratios of LIBRAvI, LIBRAvII, and LIBRAvIII was 43-, 22-, and 28-fold, respectively. This fluorescence-based method further showed that several ADA analogs had significant differences with respect to subtype selectivity and potency. These results highlight the important role played by the O-glycosidic structure of ADA in the selectivity of the ligands for IP3R1, as evidenced by the modified selectivity following replacement of the 5′-hydroxyl with a phenyl or phenethyl group. We also found that one ADA analog 5′-deoxy-5′-phenyladenophostin A possessed a partial agonistic effect on IP3R1. Together, the novel fluorescent methods described herein are useful for the evaluation of properties of IP3R ligands, including potency, efficacy, and subtype selectivity.

Keywords
ADA, adenophostin A; IP3, inositol 1,4,5-trisphosphate; IP3R, inositol 1,4,5-trisphosphate receptor; FRET, fluorescence resonance energy transfer; LBD, ligand-binding domain of IP3R; ICM, intracellular-like medium; ECFP, enhanced cyan fluorescent protein;
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A fluorescence-based method for evaluating inositol 1,4,5-trisphosphate receptor ligands: Determination of subtype selectivity and partial agonist effects
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 167, Issue 3, 10 September 2013, Pages 248–254
Authors
, , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us