Characterization of a ryanodine receptor in Periplaneta americana

J Recept Signal Transduct Res. 1997 Jan-May;17(1-3):185-97. doi: 10.3109/10799899709036603.

Abstract

Specific binding sites for the alkaloid ryanodine were characterized in membrane preparations from sarcoplasmatic reticulum of Periplaneta americana skeletal muscle. Binding of [3H]ryanodine was optimal at pH 8 and at CaCl2 concentration of about 300 mumol l-1. The Ca-chelating agents EGTA (100 mumol l-1) and EDTA (100 mumol l-1) abolished 95% and 90% of the [3H]ryanodine binding respectively. Preincubation with Ca2+ (100 mumol l-1) restored the ryanodine binding in presence of up to 300 mumol l-1 EGTA. Radioligand binding experiments showed one class of high affinity binding sites for ryanodine. Determination of rate constants revealed 7.05 x 10(6) l mol-1 min-1 for associating and 3.77 x 10(-3) min-1 for the dissociating [3H]ryanodine ryanodine receptor complex. Solubility of the ryanodine receptor was examined with different anionic, non-ionic and zwitterionic detergents. Best solubilization results of "calcium release channel" of cockroach muscle membrane preparations were obtained with the detergent CHAPS in a concentration of 5 mg ml-1.

MeSH terms

  • Animals
  • Calcium / pharmacology
  • Calcium Channels / chemistry*
  • Calcium Channels / metabolism
  • Calmodulin-Binding Proteins / chemistry*
  • Calmodulin-Binding Proteins / metabolism
  • Edetic Acid / pharmacology
  • Egtazic Acid / pharmacology
  • Female
  • Hydrogen-Ion Concentration
  • Male
  • Muscle Proteins / chemistry*
  • Muscle Proteins / metabolism
  • Muscles / drug effects
  • Muscles / metabolism
  • Periplaneta / chemistry*
  • Potassium / metabolism
  • Ryanodine / metabolism
  • Ryanodine Receptor Calcium Release Channel
  • Sodium / metabolism

Substances

  • Calcium Channels
  • Calmodulin-Binding Proteins
  • Muscle Proteins
  • Ryanodine Receptor Calcium Release Channel
  • Ryanodine
  • Egtazic Acid
  • Edetic Acid
  • Sodium
  • Potassium
  • Calcium