<i>CLCN1</i> Mutations in Czech Patients with Myotonia Congenita, <i>In Silico</i> Analysis of Novel and Known Mutations in the Human Dimeric Skeletal Muscle Chloride Channel

<div><p>Myotonia congenita (MC) is a genetic disease caused by mutations in the skeletal muscle chloride channel gene (<i>CLCN1</i>) encoding the skeletal muscle chloride channel (ClC-1). Mutations of <i>CLCN1</i> result in either autosomal dominant MC (Thomsen disease) or autosomal recessive MC (Becker disease). The ClC-1 protein is a homodimer with a separate ion pore within each monomer. Mutations causing recessive myotonia most likely affect properties of only the mutant monomer in the heterodimer, leaving the wild type monomer unaffected, while mutations causing dominant myotonia affect properties of both subunits in the heterodimer. Our study addresses two points: 1) molecular genetic diagnostics of MC by analysis of the <i>CLCN1</i> gene and 2) structural analysis of mutations in the homology model of the human dimeric ClC-1 protein. In the first part, 34 different types of <i>CLCN1</i> mutations were identified in 51 MC probands (14 mutations were new). In the second part, on the basis of the homology model we identified the amino acids which forming the dimer interface and those which form the Cl^- ion pathway. In the literature, we searched for mutations of these amino acids for which functional analyses were performed to assess the correlation between localisation of a mutation and occurrence of a dominant-negative effect (corresponding to dominant MC). This revealed that both types of mutations, with and without a dominant-negative effect, are localised at the dimer interface while solely mutations without a dominant-negative effect occur inside the chloride channel. This work is complemented by structural analysis of the homology model which provides elucidation of the effects of mutations, including a description of impacts of newly detected missense mutations. </p> </div

3D structure of the homology model of the human ClC-1 dimer (transmembrane domains are in blue and gray) with the Cl^- channel (in green) visualized in one subunit.

<p>A shows highlighted interface and the channel with the key amino acid Glu 232 (red surface) and two Cl^- ions (yellow balls) which were superimposed from the original X-ray structure of the CmClC transporter. B shows 6 new missense mutations mapped in one subunit. </p