Fluorometric assay of integrin activity with a small-molecular probe that senses the binding site microenvironment.
We have developed a small-molecular probe, consisting of cyclic RGD pentapeptides bearing a nitrobenzoxadiazole fluorophore at the 4'-residue, which detects integrin αVβ3 activity in terms of fluorescence intensity decrease due to quenching of the fluorophore by interaction with tyrosine-122 at the binding site of the protein. This probe appears to be suitable for a practical, high-throughput fluorescence assay to screen small-molecular modulators of integrin activity.
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, Japan.
Toru
T
Komatsu
Aoi
A
Takeda
Kenjiro
K
Hanaoka
Takuya
T
Terai
Tasuku
T
Ueno
Yukio
Y
Tada
Tetsuo
T
Nagano
Yasuteru
Y
Urano
D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry; D001665:Binding Sites; D020134:Catalytic Domain; D005456:Fluorescent Dyes / Q000737:chemistry; D039302:Integrin alphaVbeta3 / Q000737:chemistry / Q000378:metabolism*; D056004:Molecular Dynamics Simulation; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010456:Peptides, Cyclic / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D039302:Integrin alphaVbeta3; D015335:Molecular Probes; D009842:Oligopeptides; D010456:Peptides, Cyclic; C507169:cyclic arginine-glycine-aspartic acid peptide; D009327:4-Chloro-7-nitrobenzofurazan