Biochemical characterization of plasma membrane isolated from human skeletal muscle

FEBS Lett. 1985 Sep 2;188(2):222-6. doi: 10.1016/0014-5793(85)80376-8.

Abstract

Specific components of ion translocation systems were studied in excitable plasma membranes isolated from normal human muscle. Na+-K+ ATPase and ouabain-sensitive K+ phosphatase activities were 8.9 +/- 1 mumol Pi/h per mg protein and 96 +/- 9 nmol/min per mg protein, respectively. Scatchard analysis of equilibrium binding assays with [3H]ouabain showed non-linear curves consistent with high- and low-affinity sites (estimated Kd 3 nM and 0.22 microM). Two families of receptors with different affinities for a tritiated TTX derivative (estimated Kd 0.4 and 4 nM) were also identified suggesting the existence in human muscle of at least two classes of voltage-dependent Na+ channels. In addition (+)-[methyl-3H]PN200-110, a potent Ca2+ antagonist used for labeling voltage-dependent Ca2+ channels, was observed to bind to a homogeneous population of receptors in the plasma membrane (Kd = 0.2 nM).

MeSH terms

  • Calcium Channel Blockers / metabolism
  • Carrier Proteins / metabolism
  • Cell Fractionation
  • Cell Membrane / enzymology
  • Cell Membrane / metabolism
  • Enzyme Activation / drug effects
  • Humans
  • Ion Channels / metabolism
  • Isradipine
  • Membrane Lipids / analysis
  • Membrane Proteins / analysis
  • Muscles / enzymology*
  • Muscles / metabolism
  • Ouabain / pharmacology
  • Oxadiazoles / metabolism
  • Phosphoric Monoester Hydrolases / metabolism
  • Receptors, Drug / metabolism
  • Sodium Channels*
  • Sodium-Potassium-Exchanging ATPase / metabolism

Substances

  • Calcium Channel Blockers
  • Carrier Proteins
  • Ion Channels
  • Membrane Lipids
  • Membrane Proteins
  • Oxadiazoles
  • Receptors, Drug
  • Sodium Channels
  • cardiac glycoside receptors
  • tetrodotoxin-binding protein
  • Ouabain
  • Phosphoric Monoester Hydrolases
  • Sodium-Potassium-Exchanging ATPase
  • Isradipine