Skip to main content

Antiidiotypic antibody recognizes an amiloride binding domain within the alpha subunit of the epithelial Na+ channel.

Publication ,  Journal Article
Kieber-Emmons, T; Lin, C; Foster, MH; Kleyman, TR
Published in: J Biol Chem
April 2, 1999

We previously raised an antibody (RA6.3) by an antiidiotypic approach which was designed to be directed against an amiloride binding domain on the epithelial Na+ channel (ENaC). This antibody mimicked amiloride in that it inhibited transepithelial Na+ transport across A6 cell monolayers. RA6.3 recognized a 72-kDa polypeptide in A6 epithelia treated with tunicamycin, consistent with the size of nonglycosylated Xenopus laevis alphaENaC. RA6.3 specifically recognized an amiloride binding domain within the alpha-subunit of mouse and bovine ENaC. The deduced amino acid sequence of RA6.3 was used to generate a three-dimensional model structure of the antibody. The combining site of RA6.3 was epitope mapped using a novel computer-based strategy. Organic residues that potentially interact with the RA6.3 combining site were identified by data base screening using the program LUDI. Selected residues docked to the antibody in a manner corresponding to the ordered linear array of amino acid residues within an amiloride binding domain on the alpha-subunit of ENaC. A synthetic peptide spanning this domain inhibited the binding of RA6.3 to alphaENaC. This analysis provided a novel approach to develop models of antibody-antigen interaction as well as a molecular perspective of RA6.3 binding to an amiloride binding domain within alphaENaC.

Duke Scholars

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

April 2, 1999

Volume

274

Issue

14

Start / End Page

9648 / 9655

Location

United States

Related Subject Headings

  • Xenopus laevis
  • Sodium Channels
  • Protein Conformation
  • Molecular Weight
  • Molecular Sequence Data
  • Models, Molecular
  • Mice
  • Epithelial Sodium Channels
  • DNA, Complementary
  • Computer Simulation
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Kieber-Emmons, T., Lin, C., Foster, M. H., & Kleyman, T. R. (1999). Antiidiotypic antibody recognizes an amiloride binding domain within the alpha subunit of the epithelial Na+ channel. J Biol Chem, 274(14), 9648–9655. https://doi.org/10.1074/jbc.274.14.9648
Kieber-Emmons, T., C. Lin, M. H. Foster, and T. R. Kleyman. “Antiidiotypic antibody recognizes an amiloride binding domain within the alpha subunit of the epithelial Na+ channel.J Biol Chem 274, no. 14 (April 2, 1999): 9648–55. https://doi.org/10.1074/jbc.274.14.9648.
Kieber-Emmons T, Lin C, Foster MH, Kleyman TR. Antiidiotypic antibody recognizes an amiloride binding domain within the alpha subunit of the epithelial Na+ channel. J Biol Chem. 1999 Apr 2;274(14):9648–55.
Kieber-Emmons, T., et al. “Antiidiotypic antibody recognizes an amiloride binding domain within the alpha subunit of the epithelial Na+ channel.J Biol Chem, vol. 274, no. 14, Apr. 1999, pp. 9648–55. Pubmed, doi:10.1074/jbc.274.14.9648.
Kieber-Emmons T, Lin C, Foster MH, Kleyman TR. Antiidiotypic antibody recognizes an amiloride binding domain within the alpha subunit of the epithelial Na+ channel. J Biol Chem. 1999 Apr 2;274(14):9648–9655.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

April 2, 1999

Volume

274

Issue

14

Start / End Page

9648 / 9655

Location

United States

Related Subject Headings

  • Xenopus laevis
  • Sodium Channels
  • Protein Conformation
  • Molecular Weight
  • Molecular Sequence Data
  • Models, Molecular
  • Mice
  • Epithelial Sodium Channels
  • DNA, Complementary
  • Computer Simulation