Ryanodine receptors (RyRs) are ion channels that regulate muscle contraction by releasing calcium ions from intracellular stores into the cytoplasm. Mutations in skeletal muscle RyR (RyR1) give rise to congenital diseases such as central core disease. The absence of high-resolution structures of RyR1 has limited our understanding of channel function and disease mechanisms at the molecular level. Here, we report a structural model of the pore-forming region of RyR1. Molecular dynamics simulations show high ion binding to putative pore residues D4899, E4900, D4938, and D4945, which are experimentally known to be critical for channel conductance and selectivity. We also observe preferential localization of Ca2+ over K+ in the selectivity filter of RyR1. Simulations of RyR1-D4899Q mutant show a loss of preference to Ca2+ in the selectivity filter as seen experimentally. Electrophysiological experiments on a central core disease mutant, RyR1-G4898R, show constitutively open channels that conduct K+ but not Ca2+. Our simulations with G4898R likewise show a decrease in the preference of Ca2+ over K+ in the selectivity filter. Together, the computational and experimental results shed light on ion conductance and selectivity of RyR1 at an atomistic level.
Ryanodine receptors (RyRs) are ion channels present in the membranes of an intracellular calcium storage organelle, the sarcoplasmic reticulum. Nerve impulse triggers the opening of RyR channels, thus increasing the cytoplasmic calcium levels, which subsequently leads to muscle contraction. Congenital mutations in a specific type of RyR that is present in skeletal muscles, RyR1, lead to central core disease (CCD), which leads to weakened muscle. RyR1 mutations also render patients to be highly susceptible to malignant hyperthermia, an adverse reaction to general anesthesia. Although it is generally known that CCD mutations abort RyR1 function, the molecular basis of RyR1 dysfunction remains largely unknown because of the lack of atomic-level structure. Here, we present a structural model of the RyR1 pore region, where many of the CCD mutations are located. Molecular dynamics simulations of the pore region confirm the positions of residues experimentally known to be relevant for function. Furthermore, electrophysiological experiments and simulations shed light on the loss of function of CCD mutant channels. The combined theoretical and experimental studies on RyR1 elucidate the ion conduction pathway of RyR1 and a potential molecular origin of muscle diseases.