Determination of Binding Constants of Polyethylene Glycol Vancomycin Derivatives to Peptide Ligands Using Affinity Capillary Electrophoresis
نویسندگان
چکیده
Vancomycin (Van) from Streptomyces orientalis has been derivatized with polyethylene glycol [PEG; PEG-550 (1), 750 (2), 1,100 (3), 2,000 (4), 5,000 (5), and 8,000 (6) g mol] at the Nterminus of the glycopeptide backbone and their binding to D-Ala-D-Ala terminus peptides assessed using affinity capillary electrophoresis (ACE). Utilizing ACE, a plug of Van-PEG and non-interacting standards are injected and electrophoresed. Analysis of the change in the relative migration time ratio of the Van-PEG species, relative to the non-interacting standards, as a function of the concentration of peptide, yields a value for the binding constant (Kb). Values of Kb for N-acetyl-D-Ala-D-Ala, 7 to the Van-PEG derivatives are weaker than those for Na,Nediacetyl-Lys-D-Ala-D-Ala, 8 (for example, values of Kb for 7-1 and 8-1 are 1.8 and 47.7 · 10 M, respectively). These results demonstrate that derivatization of Van with PEG has little effect on the affinity of D-Ala-D-Ala peptide ligands to it. The findings further prove the versatility of ACE and its ability to estimate binding parameters of ligands to antibiotics.
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