A central core disease mutation in the Ca2+-binding site of skeletal muscle ryanodine receptor impairs single-channel regulation

Venkat R. Chirasani, Le Xu, Hannah G. Addis, Daniel A. Pasek, Nikolay Dokholyan, Gerhard Meissner, Naohiro Yamaguchi

Research output: Contribution to journalArticle

Abstract

Cryoelectron microscopy and mutational analyses have shown that type 1 ryanodine receptor (RyR1) amino acid residues RyR1-E3893,-E3967, and-T5001 are critical for Ca2+-mediated activation of skeletal muscle Ca2+ release channel. De novo missense mutation RyR1-Q3970K in the secondary binding sphere of Ca2+ was reported in association with central core disease (CCD) in a 2-yr-old boy. Here, we characterized recombinant RyR1-Q3970K mutant by cellular Ca2+ release measurements, single-channel recordings, and computational methods. Caffeine-induced Ca2+ release studies indicated that RyR1-Q3970K formed caffeine-sensitive, Ca2+-con-ducting channel in HEK293 cells. However, in single-channel recordings, RyR1-Q3970K displayed low Ca2+-dependent channel activity and greatly reduced activation by caffeine or ATP. A RyR1-Q3970E mutant corresponds to missense mutation RyR2-Q3925E associated with arrhythmogenic syndrome in cardiac muscle. RyR1-Q3970E also formed caffeine-induced Ca2+ release in HEK293 cells and exhibited low activity in the presence of the activating ligand Ca2+ but, in contrast to RyR1-Q3970K, was activated by ATP and caffeine in single-channel recordings. Computational analyses suggested distinct structural rearrangements in the secondary binding sphere of Ca2+ of the two mutants, whereas the interaction of Ca2+ with directly interacting RyR1 amino acid residues Glu3893, Glu3967, and Thr5001 was only minimally affected. We conclude that RyR1-Q3970 has a critical role in Ca2+-dependent activation of RyR1 and that a missense RyR1-Q3970K mutant may give rise to myopathy in skeletal muscle.

Original languageEnglish (US)
Pages (from-to)C358-C365
JournalAmerican Journal of Physiology - Cell Physiology
Volume317
Issue number2
DOIs
StatePublished - Aug 1 2019

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Central Core Myopathy
Ryanodine Receptor Calcium Release Channel
Skeletal Muscle
Binding Sites
Mutation
Caffeine
HEK293 Cells
Missense Mutation
Adenosine Triphosphate
Amino Acid Receptors
Cryoelectron Microscopy

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cell Biology

Cite this

Chirasani, Venkat R. ; Xu, Le ; Addis, Hannah G. ; Pasek, Daniel A. ; Dokholyan, Nikolay ; Meissner, Gerhard ; Yamaguchi, Naohiro. / A central core disease mutation in the Ca2+-binding site of skeletal muscle ryanodine receptor impairs single-channel regulation. In: American Journal of Physiology - Cell Physiology. 2019 ; Vol. 317, No. 2. pp. C358-C365.
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A central core disease mutation in the Ca2+-binding site of skeletal muscle ryanodine receptor impairs single-channel regulation. / Chirasani, Venkat R.; Xu, Le; Addis, Hannah G.; Pasek, Daniel A.; Dokholyan, Nikolay; Meissner, Gerhard; Yamaguchi, Naohiro.

In: American Journal of Physiology - Cell Physiology, Vol. 317, No. 2, 01.08.2019, p. C358-C365.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A central core disease mutation in the Ca2+-binding site of skeletal muscle ryanodine receptor impairs single-channel regulation

AU - Chirasani, Venkat R.

AU - Xu, Le

AU - Addis, Hannah G.

AU - Pasek, Daniel A.

AU - Dokholyan, Nikolay

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AU - Yamaguchi, Naohiro

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AB - Cryoelectron microscopy and mutational analyses have shown that type 1 ryanodine receptor (RyR1) amino acid residues RyR1-E3893,-E3967, and-T5001 are critical for Ca2+-mediated activation of skeletal muscle Ca2+ release channel. De novo missense mutation RyR1-Q3970K in the secondary binding sphere of Ca2+ was reported in association with central core disease (CCD) in a 2-yr-old boy. Here, we characterized recombinant RyR1-Q3970K mutant by cellular Ca2+ release measurements, single-channel recordings, and computational methods. Caffeine-induced Ca2+ release studies indicated that RyR1-Q3970K formed caffeine-sensitive, Ca2+-con-ducting channel in HEK293 cells. However, in single-channel recordings, RyR1-Q3970K displayed low Ca2+-dependent channel activity and greatly reduced activation by caffeine or ATP. A RyR1-Q3970E mutant corresponds to missense mutation RyR2-Q3925E associated with arrhythmogenic syndrome in cardiac muscle. RyR1-Q3970E also formed caffeine-induced Ca2+ release in HEK293 cells and exhibited low activity in the presence of the activating ligand Ca2+ but, in contrast to RyR1-Q3970K, was activated by ATP and caffeine in single-channel recordings. Computational analyses suggested distinct structural rearrangements in the secondary binding sphere of Ca2+ of the two mutants, whereas the interaction of Ca2+ with directly interacting RyR1 amino acid residues Glu3893, Glu3967, and Thr5001 was only minimally affected. We conclude that RyR1-Q3970 has a critical role in Ca2+-dependent activation of RyR1 and that a missense RyR1-Q3970K mutant may give rise to myopathy in skeletal muscle.

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