A missense mutation converts the Na+,K+-ATPase into an ion channel and causes therapy-resistant epilepsy

• Published in: The Journal of biological chemistry Vol. 297; no. 6; p. 101355
• Main Authors: Ygberg, Sofia, Akkuratov, Evgeny E., Howard, Rebecca J., Taylan, Fulya, Jans, Daniel C., Mahato, Dhani R., Katz, Adriana, Kinoshita, Paula F., Portal, Benjamin, Nennesmo, Inger, Lindskog, Maria, Karlish, Steven J.D., Andersson, Magnus, Lindstrand, Anna, Brismar, Hjalmar, Aperia, Anita
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2021; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Anticonvulsants - pharmacology; arginine mutation; Brain - drug effects; Brain - metabolism; Brain - pathology; Cells, Cultured; de novo mutation; Drug Resistance; epilepsy; Epilepsy - drug therapy; Epilepsy - genetics; Epilepsy - pathology; Humans; Infant; K-ATPase; leak channel; Medicin och hälsovetenskap; Molecular Dynamics Simulation; Mutation, Missense - drug effects; Na,K-ATPase; Protein Subunits - analysis; Protein Subunits - genetics; Sodium-Potassium-Exchanging ATPase - analysis; Sodium-Potassium-Exchanging ATPase - genetics; Xenopus; epilepsy; CADD; WGS; OR; OS; MRI; leak channel; GFP; de novo mutation; ExAC; MAP2; Na,K-ATPase; SNV; arginine mutation; DOPC
• ISSN: 0021-9258; 1083-351X; 1083-351X
• DOI: 10.1016/j.jbc.2021.101355

The ion pump Na+,K+-ATPase is a critical determinant of neuronal excitability; however, its role in the etiology of diseases of the central nervous system (CNS) is largely unknown. We describe here the molecular phenotype of a Trp931Arg mutation of the Na+,K+-ATPase catalytic α1 subunit in an infant diagnosed with therapy-resistant lethal epilepsy. In addition to the pathological CNS phenotype, we also detected renal wasting of Mg2+. We found that membrane expression of the mutant α1 protein was low, and ion pumping activity was lost. Arginine insertion into membrane proteins can generate water-filled pores in the plasma membrane, and our molecular dynamic (MD) simulations of the principle states of Na+,K+-ATPase transport demonstrated massive water inflow into mutant α1 and destabilization of the ion-binding sites. MD simulations also indicated that a water pathway was created between the mutant arginine residue and the cytoplasm, and analysis of oocytes expressing mutant α1 detected a nonspecific cation current. Finally, neurons expressing mutant α1 were observed to be depolarized compared with neurons expressing wild-type protein, compatible with a lowered threshold for epileptic seizures. The results imply that Na+,K+-ATPase should be considered a neuronal locus minoris resistentia in diseases associated with epilepsy and with loss of plasma membrane integrity.