Partial-filling affinity capillary electrophoresis.
Partial-filling affinity capillary electrophoresis (PFACE) is used to examine the binding interactions between two model biological systems: D-Ala-D-Ala terminus peptides to the glycopeptide antibiotic vancomycin (Van) from Streptomyces orientalis, and arylsulfonamides to carbonic anhydrase B (CAB, EC 4.2.1.1, bovine erythrocytes). Using these two systems, modifications in the PFACE technique are demonstrated including flow-through PFACE (FTPFACE), competitive flow-through PFACE (CFTPFACE), on-column ligand synthesis PFACE (OCLSPFACE), and multiple-step ligand injection PFACE (MSLIPFACE). In PFACE small plugs of sample are injected into the capillary column and an equilibrium is established between receptor and ligand during electrophoresis. Binding constants are then obtained by Scatchard analysis using changes in the migration time of the receptor/ligand on changing the concentration of the ligand/receptor. Data demonstrating the quantitative potential of these methods are presented. This review focuses on the unique capabilities of the different PFACE techniques as applied to two model biological systems.
Villareal
Valerie
V
Department of Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, CA 90032-8202, USA.
Kaddis
John
J
Azad
Maryam
M
Zurita
Cecilia
C
Silva
Isba
I
Hernandez
Lili
L
Rudolph
Marcellus
M
Moran
Julio
J
Gomez
Frank A
FA
D024401:Carbonic Anhydrase I / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D007700:Kinetics; D008024:Ligands; D010455:Peptides / Q000032:analysis / Q000378:metabolism; D011485:Protein Binding; D011955:Receptors, Drug / Q000032:analysis / Q000378:metabolism; D014640:Vancomycin / Q000378:metabolism*
D008024:Ligands; D010455:Peptides; D011955:Receptors, Drug; D014640:Vancomycin; D024401:Carbonic Anhydrase I