title: Investigations of the contribution of a putative glycine hinge to ryanodine receptor channel gating.
abstract: Ryanodine receptor channels (RyR) are key components of striated muscle excitation-contraction coupling, and alterations in their function underlie both inherited and acquired disease. A full understanding of the disease process will require a detailed knowledge of the mechanisms and structures involved in RyR function. Unfortunately, high-resolution structural data, such as exist for K(+)-selective channels, are not available for RyR. In the absence of these data, we have used modeling to identify similarities in the structural elements of K(+) channel pore-forming regions and postulated equivalent regions of RyR. This has identified a sequence of residues in the cytosolic cavity-lining transmembrane helix of RyR (G(4864)LIIDA(4869) in RyR2) analogous to the glycine hinge motif present in many K(+) channels. Gating in these K(+) channels can be disrupted by substitution of residues for the hinge glycine. We investigated the involvement of glycine 4864 in RyR2 gating by monitoring properties of recombinant human RyR2 channels in which this glycine is replaced by residues that alter gating in K(+) channels. Our data demonstrate that introducing alanine at position 4864 produces no significant change in RyR2 function. In contrast, function is altered when glycine 4864 is replaced by either valine or proline, the former preventing channel opening and the latter modifying both ion translocation and gating. Our studies reveal novel information on the structural basis of RyR gating, identifying both similarities with, and differences from, K(+) channels. Glycine 4864 is not absolutely required for channel gating, but some flexibility at this point in the cavity-lining transmembrane helix is necessary for normal RyR function.
authors: Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4XN, Wales, United Kingdom.
authors: Joanne
authors: J
authors: Euden
authors: Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4XN, Wales, United Kingdom.
authors: Sammy A
authors: SA
authors: Mason
authors: Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4XN, Wales, United Kingdom.
authors: Cedric
authors: C
authors: Viero
authors: Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4XN, Wales, United Kingdom.
authors: N Lowri
authors: NL
authors: Thomas
authors: Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4XN, Wales, United Kingdom. Electronic address: williamsaj9@cardiff.ac.uk.
authors: Alan J
authors: AJ
authors: Williams
mesh_terms: D019943:Amino Acid Substitution; D018360:Crystallography, X-Ray; D005998:Glycine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology*; D017136:Ion Transport / Q000502:physiology; D020125:Mutation, Missense; D017434:Protein Structure, Tertiary; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D019837:Ryanodine Receptor Calcium Release Channel; D005998:Glycine
keywords: Calcium Channels; Calcium Intracellular Release; Glycine Hinge; Ion Channels; Ryanodine Receptor; Sarcoplasmic Reticulum (SR); Single Channel Gating
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