The calmodulin regulator protein, PEP-19, sensitizes ATP-induced Ca 2+ release

Xu Wang, Liang Wen Xiong, Amina El Ayadi, Darren Boehning, John A. Putkey

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: PEP-19 modulates the kinetics of Ca2+ binding to CaM. Results: An acidic region in PEP-19 binds Ca2+ and is essential for both modulating Ca2+ binding to CaM and sensitizing cells to ATP-induced Ca2+ release. Conclusion: Simply binding to CaM is not sufficient to account for the biological activities of PEP-19. Significance: Regulating ligand-induced Ca2+ release gives PEP-19 the potential to broadly affect cell signaling. PEP-19 is a small, intrinsically disordered protein that binds to the C-domain of calmodulin (CaM) via an IQ motif and tunes its Ca2+ binding properties via an acidic sequence. We show here that the acidic sequence of PEP-19 has intrinsic Ca2+ binding activity, which may modulate Ca2+ binding to CaM by stabilizing an initial Ca2+-CaM complex or by electrostatically steering Ca 2+ to and from CaM. Because PEP-19 is expressed in cells that exhibit highly active Ca2+ dynamics, we tested the hypothesis that it influences ligand-dependent Ca2+release.We show that PEP-19 increases the sensitivity of HeLa cells to ATPinduced Ca2+ release to greatly increase the percentage of cells responding to sub-saturating doses of ATP and increases the frequency of Ca2+ oscillations. Mutations in the acidic sequence of PEP-19 that inhibit or prevent it from modulating Ca2+ binding to CaM greatly inhibit its effect on ATP-induced Ca2+ release. Thus, this cellular effect of PEP-19 does not depend simply on binding toCaMvia the IQ motif but requires its acidic metal binding domain. Tuning the activities of Ca2+ mobilization pathways places PEP-19 at the top of CaM signaling cascades, with great potential to exert broad effects on downstream CaM targets, thus expanding the biological significance of this small regulator of CaM signaling.

Original languageEnglish (US)
Pages (from-to)2040-2048
Number of pages9
JournalJournal of Biological Chemistry
Volume288
Issue number3
DOIs
StatePublished - Jan 18 2013

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Calmodulin
Adenosine Triphosphate
Proteins
Peptamen
Intrinsically Disordered Proteins
Cell signaling
Ligands
Bioactivity
HeLa Cells
Tuning
Metals
Mutation
Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The calmodulin regulator protein, PEP-19, sensitizes ATP-induced Ca 2+ release. / Wang, Xu; Xiong, Liang Wen; Ayadi, Amina El; Boehning, Darren; Putkey, John A.

In: Journal of Biological Chemistry, Vol. 288, No. 3, 18.01.2013, p. 2040-2048.

Research output: Contribution to journalArticle

Wang, Xu ; Xiong, Liang Wen ; Ayadi, Amina El ; Boehning, Darren ; Putkey, John A. / The calmodulin regulator protein, PEP-19, sensitizes ATP-induced Ca 2+ release. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 3. pp. 2040-2048.
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abstract = "Background: PEP-19 modulates the kinetics of Ca2+ binding to CaM. Results: An acidic region in PEP-19 binds Ca2+ and is essential for both modulating Ca2+ binding to CaM and sensitizing cells to ATP-induced Ca2+ release. Conclusion: Simply binding to CaM is not sufficient to account for the biological activities of PEP-19. Significance: Regulating ligand-induced Ca2+ release gives PEP-19 the potential to broadly affect cell signaling. PEP-19 is a small, intrinsically disordered protein that binds to the C-domain of calmodulin (CaM) via an IQ motif and tunes its Ca2+ binding properties via an acidic sequence. We show here that the acidic sequence of PEP-19 has intrinsic Ca2+ binding activity, which may modulate Ca2+ binding to CaM by stabilizing an initial Ca2+-CaM complex or by electrostatically steering Ca 2+ to and from CaM. Because PEP-19 is expressed in cells that exhibit highly active Ca2+ dynamics, we tested the hypothesis that it influences ligand-dependent Ca2+release.We show that PEP-19 increases the sensitivity of HeLa cells to ATPinduced Ca2+ release to greatly increase the percentage of cells responding to sub-saturating doses of ATP and increases the frequency of Ca2+ oscillations. Mutations in the acidic sequence of PEP-19 that inhibit or prevent it from modulating Ca2+ binding to CaM greatly inhibit its effect on ATP-induced Ca2+ release. Thus, this cellular effect of PEP-19 does not depend simply on binding toCaMvia the IQ motif but requires its acidic metal binding domain. Tuning the activities of Ca2+ mobilization pathways places PEP-19 at the top of CaM signaling cascades, with great potential to exert broad effects on downstream CaM targets, thus expanding the biological significance of this small regulator of CaM signaling.",
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AB - Background: PEP-19 modulates the kinetics of Ca2+ binding to CaM. Results: An acidic region in PEP-19 binds Ca2+ and is essential for both modulating Ca2+ binding to CaM and sensitizing cells to ATP-induced Ca2+ release. Conclusion: Simply binding to CaM is not sufficient to account for the biological activities of PEP-19. Significance: Regulating ligand-induced Ca2+ release gives PEP-19 the potential to broadly affect cell signaling. PEP-19 is a small, intrinsically disordered protein that binds to the C-domain of calmodulin (CaM) via an IQ motif and tunes its Ca2+ binding properties via an acidic sequence. We show here that the acidic sequence of PEP-19 has intrinsic Ca2+ binding activity, which may modulate Ca2+ binding to CaM by stabilizing an initial Ca2+-CaM complex or by electrostatically steering Ca 2+ to and from CaM. Because PEP-19 is expressed in cells that exhibit highly active Ca2+ dynamics, we tested the hypothesis that it influences ligand-dependent Ca2+release.We show that PEP-19 increases the sensitivity of HeLa cells to ATPinduced Ca2+ release to greatly increase the percentage of cells responding to sub-saturating doses of ATP and increases the frequency of Ca2+ oscillations. Mutations in the acidic sequence of PEP-19 that inhibit or prevent it from modulating Ca2+ binding to CaM greatly inhibit its effect on ATP-induced Ca2+ release. Thus, this cellular effect of PEP-19 does not depend simply on binding toCaMvia the IQ motif but requires its acidic metal binding domain. Tuning the activities of Ca2+ mobilization pathways places PEP-19 at the top of CaM signaling cascades, with great potential to exert broad effects on downstream CaM targets, thus expanding the biological significance of this small regulator of CaM signaling.

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