title: Using affinity capillary electrophoresis to study the interaction of the extracellular binding domain of erythropoietin receptor with peptides.
abstract: We have shown that affinity capillary electrophoresis (ACE) can be utilized to screen peptides that bind to the extracellular binding domain of the erythropoietin receptor (EBP). The comparison of the cyclic peptides GGTYSCHFGPLTWVCKPQGG (EMP1) GGTYSCHFGPLTAVCKPQGG (EMP13), and LGRKYSCHFGPLTWVCQPAKKD (EMP37) with the linear peptides HFGPLTWV (EMP26) and FMRF as ACE buffer additives were investigated. When EMP1 and EMP37 were the buffer additives, an abrupt change in the electrophoretic mobility of EBP was observed in the electropherogram. When EMP13, EMP26, and FMRF were examined under identical ACE conditions as EMP1 and EMP37, no significant change in the electrophoretic mobility of EBP was observed. These results correlate well with previously reported IC50 competitive binding data; that is, EMP1 and EMP37 bind to EBP while EMP13 and EMP26 bind very weakly. These observations strongly infer that peptide.EBP dimerization were induced by EMP1, and EMP37 but not by EMP13, EMP26 or FMRF. This ACE method provides a rapid tool for the detection of small peptides or drugs that bind to EBP.
authors.affiliation: The R.W. Johnson Pharmaceutical Research Institute, Spring House, PA, USA. gcaldwel@prius.jnj.com
authors.forename: G W
authors.initials: GW
authors.lastname: Caldwell
authors.forename: P A
authors.initials: PA
authors.lastname: McDonnell
authors.forename: J A
authors.initials: JA
authors.lastname: Masucci
authors.forename: D L
authors.initials: DL
authors.lastname: Johnson
authors.forename: L K
authors.initials: LK
authors.lastname: Jolliffe
mesh_terms: D000595:Amino Acid Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D008024:Ligands; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000737:chemistry*; D011485:Protein Binding; D017467:Receptors, Erythropoietin / Q000737:chemistry*
chemical_list: D008024:Ligands; D010456:Peptides, Cyclic; D017467:Receptors, Erythropoietin