title: Estimation of receptor-ligand interactions by the use of a two-marker system in affinity capillary electrophoresis.
abstract: The study of receptor-ligand interactions by affinity capillary electrophoresis (ACE) requires an accurate form of analysis. Here, we examine the use of two noninteracting standards (markers) in the analysis of binding constant data in ACE studies. This concept is demonstrated using two model systems: carbonic anhydrase B (CAB, EC 4.2.1.1) and arylsulfonamides, and vancomycin (Van) from Streptomyces orientalis and the dipeptide N-acetyl-d-Ala-d-Ala. In this procedure a plug of receptor and noninteracting standards is injected, and analysis of the change in the relative migration time ratio of the receptor, relative to the noninteracting standards, as a function of the concentration of the ligand yields a value for the binding constant. The findings described here demonstrate that data from ACE studies can best be analyzed using two noninteracting standards, yielding values comparable to those estimated using other binding and ACE techniques.
authors.affiliation: Department of Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, California, 90032, USA.
authors.forename: E
authors.initials: E
authors.lastname: Mito
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: S
authors.initials: S
authors.lastname: Esquivel
authors.forename: F A
authors.initials: FA
authors.lastname: Gomez
mesh_terms: D001667:Binding, Competitive; D002256:Carbonic Anhydrases / Q000378:metabolism*; D004151:Dipeptides / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000379:methods*; D007700:Kinetics; D008024:Ligands; D013302:Streptomyces; D013449:Sulfonamides / Q000378:metabolism*; D013997:Time Factors; D014640:Vancomycin / Q000378:metabolism*
chemical_list: D004151:Dipeptides; D008024:Ligands; D013449:Sulfonamides; C031735:acetylalanylalanine; D014640:Vancomycin; D002256:Carbonic Anhydrases