ClC-1 mutations in myotonia congenita patients: Insights into molecular gating mechanisms and genotype-phenotype correlation

P. Imbrici, L. Maggi, G. F. Mangiatordi, M. M. Dinardo, C. Altamura, R. Brugnoni, D. Alberga, G. Lauria Pinter, G. Ricci, G. Siciliano, R. Micheli, G. Annicchiarico, G. Lattanzi, O. Nicolotti, L. Morandi, P. Bernasconi, J. F. Desaphy, R. Mantegazza, D. Conte Camerino

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Abstract

Myotonia congenita is an inherited disease caused by loss-of-function mutations of the skeletal muscle ClC-1 chloride channel, characterized by impaired muscle relaxation after contraction and stiffness. In the present study, we provided an in-depth characterization of F484L, a mutation previously identified in dominant myotonia, in order to define the genotype-phenotype correlation, and to elucidate the contribution of this pore residue to the mechanisms of ClC-1 gating. Patch-clamp recordings showed that F484L reduced chloride currents at every tested potential and dramatically right-shifted the voltage dependence of slow gating, thus contributing to the mild clinical phenotype of affected heterozygote carriers. Unlike dominant mutations located at the dimer interface, no dominant-negative effect was observed when F484L mutant subunits were co-expressed with wild type. Molecular dynamics simulations further revealed that F484L affected the slow gate by increasing the frequency and stability of the H-bond formation between the pore residue E232 and the R helix residue Y578. In addition, using patch-clamp electrophysiology, we characterized three other myotonic ClC-1 mutations. We proved that the dominant L198P mutation in the channel pore also right-shifted the voltage dependence of slow gating, recapitulating mild myotonia. The recessive V640G mutant drastically reduced channel function, which probably accounts for myotonia. In contrast, the recessive L628P mutant produced currents very similar to wild type, suggesting that the occurrence of the compound truncating mutation (Q812X) or other muscle-specific mechanisms accounted for the severe symptoms observed in this family. Our results provide novel insight into the molecular mechanisms underlying normal and altered ClC-1 function. Journal compilation

Original languageEnglish
Pages (from-to)4181-4199
Number of pages19
JournalJournal of Physiology
Volume593
Issue number18
DOIs
Publication statusPublished - Sep 1 2015

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Myotonia Congenita
Genetic Association Studies
Myotonia
Mutation
Chloride Channels
Muscle Relaxation
Electrophysiology
Molecular Dynamics Simulation
Heterozygote
Chlorides
Skeletal Muscle
Phenotype
Muscles

ASJC Scopus subject areas

  • Physiology

Cite this

Imbrici, P., Maggi, L., Mangiatordi, G. F., Dinardo, M. M., Altamura, C., Brugnoni, R., ... Camerino, D. C. (2015). ClC-1 mutations in myotonia congenita patients: Insights into molecular gating mechanisms and genotype-phenotype correlation. Journal of Physiology, 593(18), 4181-4199. https://doi.org/10.1113/JP270358

ClC-1 mutations in myotonia congenita patients : Insights into molecular gating mechanisms and genotype-phenotype correlation. / Imbrici, P.; Maggi, L.; Mangiatordi, G. F.; Dinardo, M. M.; Altamura, C.; Brugnoni, R.; Alberga, D.; Pinter, G. Lauria; Ricci, G.; Siciliano, G.; Micheli, R.; Annicchiarico, G.; Lattanzi, G.; Nicolotti, O.; Morandi, L.; Bernasconi, P.; Desaphy, J. F.; Mantegazza, R.; Camerino, D. Conte.

In: Journal of Physiology, Vol. 593, No. 18, 01.09.2015, p. 4181-4199.

Research output: Contribution to journalArticle

Imbrici, P, Maggi, L, Mangiatordi, GF, Dinardo, MM, Altamura, C, Brugnoni, R, Alberga, D, Pinter, GL, Ricci, G, Siciliano, G, Micheli, R, Annicchiarico, G, Lattanzi, G, Nicolotti, O, Morandi, L, Bernasconi, P, Desaphy, JF, Mantegazza, R & Camerino, DC 2015, 'ClC-1 mutations in myotonia congenita patients: Insights into molecular gating mechanisms and genotype-phenotype correlation', Journal of Physiology, vol. 593, no. 18, pp. 4181-4199. https://doi.org/10.1113/JP270358
Imbrici, P. ; Maggi, L. ; Mangiatordi, G. F. ; Dinardo, M. M. ; Altamura, C. ; Brugnoni, R. ; Alberga, D. ; Pinter, G. Lauria ; Ricci, G. ; Siciliano, G. ; Micheli, R. ; Annicchiarico, G. ; Lattanzi, G. ; Nicolotti, O. ; Morandi, L. ; Bernasconi, P. ; Desaphy, J. F. ; Mantegazza, R. ; Camerino, D. Conte. / ClC-1 mutations in myotonia congenita patients : Insights into molecular gating mechanisms and genotype-phenotype correlation. In: Journal of Physiology. 2015 ; Vol. 593, No. 18. pp. 4181-4199.
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abstract = "Myotonia congenita is an inherited disease caused by loss-of-function mutations of the skeletal muscle ClC-1 chloride channel, characterized by impaired muscle relaxation after contraction and stiffness. In the present study, we provided an in-depth characterization of F484L, a mutation previously identified in dominant myotonia, in order to define the genotype-phenotype correlation, and to elucidate the contribution of this pore residue to the mechanisms of ClC-1 gating. Patch-clamp recordings showed that F484L reduced chloride currents at every tested potential and dramatically right-shifted the voltage dependence of slow gating, thus contributing to the mild clinical phenotype of affected heterozygote carriers. Unlike dominant mutations located at the dimer interface, no dominant-negative effect was observed when F484L mutant subunits were co-expressed with wild type. Molecular dynamics simulations further revealed that F484L affected the slow gate by increasing the frequency and stability of the H-bond formation between the pore residue E232 and the R helix residue Y578. In addition, using patch-clamp electrophysiology, we characterized three other myotonic ClC-1 mutations. We proved that the dominant L198P mutation in the channel pore also right-shifted the voltage dependence of slow gating, recapitulating mild myotonia. The recessive V640G mutant drastically reduced channel function, which probably accounts for myotonia. In contrast, the recessive L628P mutant produced currents very similar to wild type, suggesting that the occurrence of the compound truncating mutation (Q812X) or other muscle-specific mechanisms accounted for the severe symptoms observed in this family. Our results provide novel insight into the molecular mechanisms underlying normal and altered ClC-1 function. Journal compilation",
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AU - Dinardo, M. M.

AU - Altamura, C.

AU - Brugnoni, R.

AU - Alberga, D.

AU - Pinter, G. Lauria

AU - Ricci, G.

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AU - Lattanzi, G.

AU - Nicolotti, O.

AU - Morandi, L.

AU - Bernasconi, P.

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