FKBP12 modulation of the binding of the skeletal ryanodine receptor onto the II-III loop of the dihydropyridine receptor

Fiona M. O'Reilly, Mylène Robert, I. Jóna, C. Szegedi, Mireille Albrieux, Sandrine Geib, Michel De Waard, Michel Villaz, Michel Ronjat

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

In skeletal muscle, excitation-contraction coupling involves a functional interaction between the ryanodine receptor (RyR) and the dihydropyridine receptor (DHPR). The domain corresponding to Thr671-Leu690 of the II-III loop of the skeletal DHPR α1-subunit is able to regulate RyR properties and calcium release from sarcoplasmic reticulum, whereas the domain corresponding to Glu724-Pro760 antagonizes this effect. Two peptides, covering these sequences (peptide ASk and CSk, respectively) were immobilized on polystyrene beads. We demonstrate that peptide ASk binds to the skeletal isoform of RyR (RyR1) whereas peptide CSk does not. Using surface plasmon resonance detection, we show that 1) domain Thr671-Leu690 is the only sequence of the II-III loop binding with RyR1 and 2) the interaction of peptide Ask with RyR1 is not modulated by Ca2+ (pCa 9-2) nor by Mg2+ (up to 10 mM). In contrast, this interaction is strongly potentiated by the immunophilin FKBP12 (EC50 = 10 nM) and inhibited by both rapamycin (IC50 = 5 nM) and FK506. Peptide ASk induces a 300% increase of the opening probability of the RyR1 incorporated in lipid bilayer. Removal of FKBP12 from RyR1 completely abolishes this effect of domain ASk on RyR1 channel behavior. These results demonstrate a direct interaction of the RyR1 with the discrete domain of skeletal DHPR α1-subunit corresponding to Thr671-Leu690 and show that the association of FKBP12 with RyR1 specifically modulates this interaction.

Original languageEnglish
Pages (from-to)145-155
Number of pages11
JournalBiophysical Journal
Volume82
Issue number1
Publication statusPublished - 2002

Fingerprint

Tacrolimus Binding Protein 1A
L-Type Calcium Channels
Ryanodine Receptor Calcium Release Channel
Peptides
Immunophilins
Excitation Contraction Coupling
Surface Plasmon Resonance
Polystyrenes
Lipid Bilayers
Sarcoplasmic Reticulum
Tacrolimus
Sirolimus
Inhibitory Concentration 50

ASJC Scopus subject areas

  • Biophysics

Cite this

FKBP12 modulation of the binding of the skeletal ryanodine receptor onto the II-III loop of the dihydropyridine receptor. / O'Reilly, Fiona M.; Robert, Mylène; Jóna, I.; Szegedi, C.; Albrieux, Mireille; Geib, Sandrine; De Waard, Michel; Villaz, Michel; Ronjat, Michel.

In: Biophysical Journal, Vol. 82, No. 1, 2002, p. 145-155.

Research output: Contribution to journalArticle

O'Reilly, FM, Robert, M, Jóna, I, Szegedi, C, Albrieux, M, Geib, S, De Waard, M, Villaz, M & Ronjat, M 2002, 'FKBP12 modulation of the binding of the skeletal ryanodine receptor onto the II-III loop of the dihydropyridine receptor', Biophysical Journal, vol. 82, no. 1, pp. 145-155.
O'Reilly, Fiona M. ; Robert, Mylène ; Jóna, I. ; Szegedi, C. ; Albrieux, Mireille ; Geib, Sandrine ; De Waard, Michel ; Villaz, Michel ; Ronjat, Michel. / FKBP12 modulation of the binding of the skeletal ryanodine receptor onto the II-III loop of the dihydropyridine receptor. In: Biophysical Journal. 2002 ; Vol. 82, No. 1. pp. 145-155.
@article{9a8b16c5ba5c46ad8ed0bf6f3d02ed2d,
title = "FKBP12 modulation of the binding of the skeletal ryanodine receptor onto the II-III loop of the dihydropyridine receptor",
abstract = "In skeletal muscle, excitation-contraction coupling involves a functional interaction between the ryanodine receptor (RyR) and the dihydropyridine receptor (DHPR). The domain corresponding to Thr671-Leu690 of the II-III loop of the skeletal DHPR α1-subunit is able to regulate RyR properties and calcium release from sarcoplasmic reticulum, whereas the domain corresponding to Glu724-Pro760 antagonizes this effect. Two peptides, covering these sequences (peptide ASk and CSk, respectively) were immobilized on polystyrene beads. We demonstrate that peptide ASk binds to the skeletal isoform of RyR (RyR1) whereas peptide CSk does not. Using surface plasmon resonance detection, we show that 1) domain Thr671-Leu690 is the only sequence of the II-III loop binding with RyR1 and 2) the interaction of peptide Ask with RyR1 is not modulated by Ca2+ (pCa 9-2) nor by Mg2+ (up to 10 mM). In contrast, this interaction is strongly potentiated by the immunophilin FKBP12 (EC50 = 10 nM) and inhibited by both rapamycin (IC50 = 5 nM) and FK506. Peptide ASk induces a 300{\%} increase of the opening probability of the RyR1 incorporated in lipid bilayer. Removal of FKBP12 from RyR1 completely abolishes this effect of domain ASk on RyR1 channel behavior. These results demonstrate a direct interaction of the RyR1 with the discrete domain of skeletal DHPR α1-subunit corresponding to Thr671-Leu690 and show that the association of FKBP12 with RyR1 specifically modulates this interaction.",
author = "O'Reilly, {Fiona M.} and Myl{\`e}ne Robert and I. J{\'o}na and C. Szegedi and Mireille Albrieux and Sandrine Geib and {De Waard}, Michel and Michel Villaz and Michel Ronjat",
year = "2002",
language = "English",
volume = "82",
pages = "145--155",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "1",

}

TY - JOUR

T1 - FKBP12 modulation of the binding of the skeletal ryanodine receptor onto the II-III loop of the dihydropyridine receptor

AU - O'Reilly, Fiona M.

AU - Robert, Mylène

AU - Jóna, I.

AU - Szegedi, C.

AU - Albrieux, Mireille

AU - Geib, Sandrine

AU - De Waard, Michel

AU - Villaz, Michel

AU - Ronjat, Michel

PY - 2002

Y1 - 2002

N2 - In skeletal muscle, excitation-contraction coupling involves a functional interaction between the ryanodine receptor (RyR) and the dihydropyridine receptor (DHPR). The domain corresponding to Thr671-Leu690 of the II-III loop of the skeletal DHPR α1-subunit is able to regulate RyR properties and calcium release from sarcoplasmic reticulum, whereas the domain corresponding to Glu724-Pro760 antagonizes this effect. Two peptides, covering these sequences (peptide ASk and CSk, respectively) were immobilized on polystyrene beads. We demonstrate that peptide ASk binds to the skeletal isoform of RyR (RyR1) whereas peptide CSk does not. Using surface plasmon resonance detection, we show that 1) domain Thr671-Leu690 is the only sequence of the II-III loop binding with RyR1 and 2) the interaction of peptide Ask with RyR1 is not modulated by Ca2+ (pCa 9-2) nor by Mg2+ (up to 10 mM). In contrast, this interaction is strongly potentiated by the immunophilin FKBP12 (EC50 = 10 nM) and inhibited by both rapamycin (IC50 = 5 nM) and FK506. Peptide ASk induces a 300% increase of the opening probability of the RyR1 incorporated in lipid bilayer. Removal of FKBP12 from RyR1 completely abolishes this effect of domain ASk on RyR1 channel behavior. These results demonstrate a direct interaction of the RyR1 with the discrete domain of skeletal DHPR α1-subunit corresponding to Thr671-Leu690 and show that the association of FKBP12 with RyR1 specifically modulates this interaction.

AB - In skeletal muscle, excitation-contraction coupling involves a functional interaction between the ryanodine receptor (RyR) and the dihydropyridine receptor (DHPR). The domain corresponding to Thr671-Leu690 of the II-III loop of the skeletal DHPR α1-subunit is able to regulate RyR properties and calcium release from sarcoplasmic reticulum, whereas the domain corresponding to Glu724-Pro760 antagonizes this effect. Two peptides, covering these sequences (peptide ASk and CSk, respectively) were immobilized on polystyrene beads. We demonstrate that peptide ASk binds to the skeletal isoform of RyR (RyR1) whereas peptide CSk does not. Using surface plasmon resonance detection, we show that 1) domain Thr671-Leu690 is the only sequence of the II-III loop binding with RyR1 and 2) the interaction of peptide Ask with RyR1 is not modulated by Ca2+ (pCa 9-2) nor by Mg2+ (up to 10 mM). In contrast, this interaction is strongly potentiated by the immunophilin FKBP12 (EC50 = 10 nM) and inhibited by both rapamycin (IC50 = 5 nM) and FK506. Peptide ASk induces a 300% increase of the opening probability of the RyR1 incorporated in lipid bilayer. Removal of FKBP12 from RyR1 completely abolishes this effect of domain ASk on RyR1 channel behavior. These results demonstrate a direct interaction of the RyR1 with the discrete domain of skeletal DHPR α1-subunit corresponding to Thr671-Leu690 and show that the association of FKBP12 with RyR1 specifically modulates this interaction.

UR - http://www.scopus.com/inward/record.url?scp=0036216872&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036216872&partnerID=8YFLogxK

M3 - Article

VL - 82

SP - 145

EP - 155

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 1

ER -