Deletion of amino acids 1641-2437 from the foot region of skeletal muscle ryanodine receptor alters the conduction properties of the Ca release channel

Biophys J. 1997 Sep;73(3):1320-8. doi: 10.1016/S0006-3495(97)78165-7.

Abstract

The ryanodine receptor (RyR) of skeletal muscle contains two functional domains: a carboxyl-terminal hydrophobic domain that forms the putative conduction pore of the calcium release channel, and a large cytoplasmic domain that corresponds to the "foot structure." To understand the contribution of the foot structure to the function of the calcium release channel, we studied a RyR deletion mutant, delta(1641-2437)-RyR, in which a region that is rich in glutamate and aspartate residues (a.a. 1641-2437) was removed. The wild-type and delta(1641-2437)-RyR proteins were expressed in a Chinese hamster ovary (CHO) cell line, and functions of single calcium release channels were measured in the lipid bilayer membrane. The wild-type RyR forms functional calcium release channels with a linear current-voltage relationship similar to that of the native channel identified in the sarcoplasmic reticulum membrane of skeletal muscle, whereas the channels formed by delta(1641-2437)-RyR exhibit significant inward rectification, i.e., currents moving from cytoplasm into SR lumen were approximately 20% less than that in the opposite direction. As in to the wt-RyR channel, opening of the delta(1641-2437)-RyR channel has a bell-shaped dependence on the cytoplasmic calcium, but the calcium-dependent activation and inactivation processes of the delta(1641-2437)-RyR channel are shifted to higher calcium concentrations. Our data show that deletion of a.a. 1641-2437 from the foot region of the skeletal muscle RyR results in changes in both ion conduction and calcium-dependent regulation of the calcium release channel.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Aspartic Acid
  • CHO Cells
  • Calcium / metabolism
  • Calcium Channels / chemistry*
  • Calcium Channels / physiology*
  • Cloning, Molecular
  • Cricetinae
  • Glutamic Acid
  • Intracellular Membranes / physiology
  • Ion Channel Gating
  • Kinetics
  • Lipid Bilayers
  • Membrane Potentials
  • Microsomes / physiology
  • Muscle Proteins / chemistry*
  • Muscle Proteins / physiology*
  • Muscle, Skeletal / metabolism*
  • Mutagenesis
  • Probability
  • Rabbits
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Ryanodine Receptor Calcium Release Channel
  • Sequence Deletion
  • Transfection

Substances

  • Calcium Channels
  • Lipid Bilayers
  • Muscle Proteins
  • Recombinant Proteins
  • Ryanodine Receptor Calcium Release Channel
  • Aspartic Acid
  • Glutamic Acid
  • Calcium