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Affinity Capillary Electrophoresis to Examine Receptor-Ligand Interactions

  • Maryam Azad
  • John Kaddis
  • Valerie Villareal
  • Lili Hernandez
  • Catherine Silverio
  • Frank A. Gomez
Part of the Methods in Molecular Biology™ book series (MIMB, volume 276)

Abstract

Afffinity capillary electrophoresis (ACE) is a new analytical technique that has been shown to be an efficient and accurate tool in studying biomolecular noncovalent interactions and determining binding and dissociation constants of formed complexes. ACE uses as its basis the change in migration time of a receptor upon binding to a ligand found in the electrophoresis buffer. Subsequent Scatchard analysis using noninteracting markers realizes a binding constant. Herein, ACE and three modifications in the technique, partial-filling ACE (PFACE), flowthrough PFACE (FTPFACE), and multiple-step ligand injection ACE (MSLIACE) are used to probe the binding of ristocetin A (Rist A) and vancomycin (Van) from Streptomyces orientalis to D-Ala-D-Ala terminus peptides and carbonic anhydrase B (CAB, E.C.4.2.1.1) to arylsulfonamides.

Key Words

Affinity capillary electrophoresis binding constants carbonic anhydrase B receptor-ligand interactions ristocetin scatchard plot vancomcyin 

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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Maryam Azad
    • 1
  • John Kaddis
    • 1
  • Valerie Villareal
    • 1
  • Lili Hernandez
    • 1
  • Catherine Silverio
    • 1
  • Frank A. Gomez
    • 1
  1. 1.Department of Chemistry and BiochemistryCalifornia State UniversityLos Angeles

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