A study of ligand binding to protein using resonance energy transfer from the isoindole fluorescent label to ligand
Gryzunov, I.A.; Komarova, M.N.
Biofizika 49(6): 979-986
2004
ISSN/ISBN: 0006-3029 PMID: 15612536 Document Number: 578235
A method for studing the binging of ligands absorbing the light in the region of 350-550 nm to protein is described. The method is based on resonance energy transfer between the fluorescent label covalently bound to the protein and the ligand. The isoindole label, a product of the reaction of the protein with o-phthalaidehyde in the presence of 2-mercaptoethanol, was used as a fluorescent donor. The method was used to determine the binding parameters of a fluorescent probe (a naphthalimide derivative) with human serum albumin. A comparison of the results obtained by the resonance energy and transfer by equilibrium dialysis showed a high accuracy of the resonance energy transfer method.