Document Detail

Estimation of binding constants between ristocetin and teicoplanin and peptides using on-column ligand derivatization coupled to affinity capillary electrophoresis.
MedLine Citation:
PMID:  15042270     Owner:  NLM     Status:  MEDLINE    
This work utilizes on-column ligand synthesis and affinity capillary electrophoresis (ACE) to determine binding constants ( K(b)) of 9-flourenylmethyloxy carbonyl (Fmoc)-amino acid derivatives to the glycopeptide antibiotics ristocetin (Rist) and teicoplanin (Teic). In this technique, two separate plugs of sample are injected on to the capillary column and electrophoresed. The initial sample plug contains a D-Ala- D-Ala terminus peptide and either one or two non-interacting standard(s). The second plug contains a Fmoc-amino acid- N-hydroxysuccinimide (NHS) ester. The electrophoresis is then carried out with an increasing concentration of Rist or Teic in the running buffer. Upon electrophoresis the initial D-Ala- D-Ala peptide reacts with the Fmoc-amino acid yielding a new Fmoc-amino acid- D-Ala- D-Ala peptide derivative. Continued electrophoresis results in the binding of Rist or Teic to the Fmoc-amino acid- D-Ala- D-Ala peptide derivatives. Analysis of the change in the relative migration time ratio ( RMTR) or electrophoretic mobility (mu) of the Fmoc-amino acid- D-Ala- D-Ala peptide derivatives relative to the non-interacting standards, as a function of the concentration of Rist and Teic, yields a value for K(b). These findings demonstrate the advantage of coupling on-column ligand synthesis to ACE for estimating binding parameters between antibiotics and ligands.
Maryam Azad; Abby Brown; Isba Silva; Frank A Gomez
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.     Date:  2004-03-24
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  379     ISSN:  1618-2642     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2004 May 
Date Detail:
Created Date:  2004-04-27     Completed Date:  2004-07-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  149-55     Citation Subset:  IM    
Department of Chemistry and Biochemistry, California State University, 5151 State University Drive, Los Angeles, CA 90032-8202, USA.
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MeSH Terms
Amino Acids / chemistry
Electrophoresis, Capillary / methods
Fluorenes / chemistry
Molecular Structure
Peptides / chemistry*
Protein Binding
Ristocetin / chemistry*
Succinimides / chemistry
Teicoplanin / chemistry*
Grant Support
R15 AI055515-01/AI/NIAID NIH HHS
Reg. No./Substance:
0/Amino Acids; 0/Fluorenes; 0/Ligands; 0/N(alpha)-fluorenylmethyloxycarbonylamino acids; 0/Peptides; 0/Succinimides; 1404-55-3/Ristocetin; 6066-82-6/N-hydroxysuccinimide; 61036-62-2/Teicoplanin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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