Characterisation of peptide interactions that regulate PKCε activation

Indu R. Chandrashekaran, Raymond S. Norton, Carsten Schmitz-Peiffer

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Targeting the interaction between PKC isoforms and their anchoring proteins can specifically regulate kinase activity. εV1-2 and pseudoεRACK peptides, derived from the PKCε C2 domain, modulate its association with receptor for activated C-kinase 2 (RACK2) and thus its function. Details of these interactions remain obscure, and we therefore investigated binding of these peptides using biophysical techniques. Surface plasmon resonance (SPR) indicated that the inhibitory εV1-2 peptide bound to RACK2, and inhibited PKCε binding as expected. In contrast, SPR and NMR demonstrated that the activating pseudoεRACK peptide and related sequences did not bind to PKCε, indicating that their mechanisms of action do not involve binding to the kinase as previously proposed. Our results clarify which interactions could be targeted in developing new therapeutics that inhibit PKCε–RACK2 interaction.

Original languageEnglish
Pages (from-to)179-189
Number of pages11
JournalFEBS Letters
Volume592
Issue number2
DOIs
Publication statusPublished - 1 Jan 2018

Keywords

  • C2 domain
  • peptides
  • protein kinase C
  • RACK2
  • saturation transfer difference NMR
  • surface plasmon resonance

Cite this

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title = "Characterisation of peptide interactions that regulate PKCε activation",
abstract = "Targeting the interaction between PKC isoforms and their anchoring proteins can specifically regulate kinase activity. εV1-2 and pseudoεRACK peptides, derived from the PKCε C2 domain, modulate its association with receptor for activated C-kinase 2 (RACK2) and thus its function. Details of these interactions remain obscure, and we therefore investigated binding of these peptides using biophysical techniques. Surface plasmon resonance (SPR) indicated that the inhibitory εV1-2 peptide bound to RACK2, and inhibited PKCε binding as expected. In contrast, SPR and NMR demonstrated that the activating pseudoεRACK peptide and related sequences did not bind to PKCε, indicating that their mechanisms of action do not involve binding to the kinase as previously proposed. Our results clarify which interactions could be targeted in developing new therapeutics that inhibit PKCε–RACK2 interaction.",
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Characterisation of peptide interactions that regulate PKCε activation. / Chandrashekaran, Indu R.; Norton, Raymond S.; Schmitz-Peiffer, Carsten.

In: FEBS Letters, Vol. 592, No. 2, 01.01.2018, p. 179-189.

Research output: Contribution to journalArticleResearchpeer-review

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