[A study of ligand binding to protein using resonance energy transfer from the isoindole fluorescent label to ligand].
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-phthalaldehyde 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.
Gryzunov
Iu A
IuA
Komarova
M N
MN
D004735:Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007211:Indoles / Q000138:chemical synthesis / Q000737:chemistry*; D008024:Ligands; D008623:Mercaptoethanol / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D009764:o-Phthalaldehyde / Q000737:chemistry
D005456:Fluorescent Dyes; D007211:Indoles; D008024:Ligands; D011506:Proteins; D012709:Serum Albumin; D008623:Mercaptoethanol; D009764:o-Phthalaldehyde