Monoclonal antimyogenin antibodies define epitopes outside the bHLH domain where binding interferes with protein-protein and protein-DNA interactions

Woodring E. Wright, Ia Dac-Korytko, Karen Farmer

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

We have developed a panel of monoclonal antibodies against rat myogenin, a skeletal muscle regulatory protein of the bHLH family. Some of these monoclonals have been widely used by others, and details of their production are presented. Mapping the epitopes by immunoprecipitation of myogenin deletion mutants demonstrates that this panel recognizes epitopes spanning the entire molecule outside the HLH region. Four antibodies against epitopes outside the bHLH region interfere with the binding of myogenin/E-protein heterodimers to DNA sequences containing the myogenin heterodimer consensus recognition site. Three of these epitopes are partially masked in the heterodimers; antibody binding to these epitopes reduces the interactions between myogenin and E12. This suggests that surfaces outside the HLH dimerization domain may contribute to the stability of myogenin/E12 complexes. The binding of one antibody to its epitope did not appear to affect the myogenin/E12 interaction but nonetheless interfered with the binding of the complex to DNA, suggesting that this epitope lies near to a surface occupied by DNA.

Original languageEnglish (US)
Pages (from-to)131-138
Number of pages8
JournalDevelopmental Genetics
Volume19
Issue number2
DOIs
StatePublished - 1996

Fingerprint

Myogenin
Epitopes
Monoclonal Antibodies
DNA
Proteins
Antibodies
Epitope Mapping
Muscle Proteins
Dimerization
Immunoprecipitation
Skeletal Muscle

Keywords

  • enhancers
  • Myogenesis
  • myogenin antibodies
  • transcription factors

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology
  • Genetics

Cite this

Monoclonal antimyogenin antibodies define epitopes outside the bHLH domain where binding interferes with protein-protein and protein-DNA interactions. / Wright, Woodring E.; Dac-Korytko, Ia; Farmer, Karen.

In: Developmental Genetics, Vol. 19, No. 2, 1996, p. 131-138.

Research output: Contribution to journalArticle

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N2 - We have developed a panel of monoclonal antibodies against rat myogenin, a skeletal muscle regulatory protein of the bHLH family. Some of these monoclonals have been widely used by others, and details of their production are presented. Mapping the epitopes by immunoprecipitation of myogenin deletion mutants demonstrates that this panel recognizes epitopes spanning the entire molecule outside the HLH region. Four antibodies against epitopes outside the bHLH region interfere with the binding of myogenin/E-protein heterodimers to DNA sequences containing the myogenin heterodimer consensus recognition site. Three of these epitopes are partially masked in the heterodimers; antibody binding to these epitopes reduces the interactions between myogenin and E12. This suggests that surfaces outside the HLH dimerization domain may contribute to the stability of myogenin/E12 complexes. The binding of one antibody to its epitope did not appear to affect the myogenin/E12 interaction but nonetheless interfered with the binding of the complex to DNA, suggesting that this epitope lies near to a surface occupied by DNA.

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