Use of a two-hybrid system to investigate molecular interactions of GAP-43

Steven Chao, Larry I. Benowitz, Dimitri Krainc, Nina Irwin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

We used the 'interaction trap' (two-hybrid system) to identify polypeptides that interact with the neuronal phosphoprotein, GAP-43, in an intracellular environment. GAP-43 (neuromodulin, B-50, F1), a protein kinase C (PKC) substrate important for the growth and plasticity of neuronal connections, has been implicated in vitro in several signal transduction pathways. In the yeast-based cloning system, the only strong interaction that was detected was between GAP-43 and the calcium effector protein, calmodulin (CaM). PKC phosphorylates GAP-43 on serine 41. When we changed this serine to an aspartate residue to mimic constitutive phosphorylation, the interaction with CaM was blocked. Surprisingly, the N-terminal third of GAP-43 alone bound CaM more strongly than did intact GAP-43, suggesting that the protein's C-terminus may play a role in modulating the interaction with CaM. These results, along with other recent findings, suggest a novel role for the interaction between GAP-43 and CaM.

Original languageEnglish (US)
Pages (from-to)195-202
Number of pages8
JournalMolecular Brain Research
Volume40
Issue number2
DOIs
StatePublished - Sep 1 1996

Keywords

  • Calmodulin
  • Development
  • GAP-43
  • Growth cone
  • Phosphoprotein
  • Protein kinase C
  • Two-hybrid system

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

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