Determination of protein-ligand interactions using accelerator mass spectrometry: modified crosslinking assay.
A highly sensitive detection method for the determination of protein-ligand interactions has been developed. Radiocarbon-labeled 17beta-estradiol was incubated with estrogen receptor-alpha; as a selective binding partner, and covalently attached using crosslinking agents, to form covalently linked protein-ligand complexes. After separation using a denaturing gel, the (14)C content in the sliced gels was identified by accelerator mass spectrometry. The obtained data demonstrated specific binding of the small molecule to its binding partner. In theory, this method can be applied to most protein-ligand interaction studies.
Hah
Sang Soo
SS
Research Resource for Biomedical Accelerator Mass Spectrometry. sshah@khu.ac.kr
D002250:Carbon Radioisotopes / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004586:Electrophoresis; D004958:Estradiol / Q000737:chemistry*; D047628:Estrogen Receptor alpha / Q000737:chemistry*; D013058:Mass Spectrometry; D011485:Protein Binding; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
D002250:Carbon Radioisotopes; D003432:Cross-Linking Reagents; D047628:Estrogen Receptor alpha; D004958:Estradiol