Characterization of a ryanodine receptor in Periplaneta americana.
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.
Schmitt
M
M
Institute for Parasitology, Bayer AG, Leverkusen, Germany.
Turberg
A
A
Londershausen
M
M
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D015220:Calcium Channels / Q000737:chemistry* / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D005260:Female; D006863:Hydrogen-Ion Concentration; D008297:Male; D009124:Muscle Proteins / Q000737:chemistry* / Q000378:metabolism; D009132:Muscles / Q000187:drug effects / Q000378:metabolism; D010526:Periplaneta / Q000737:chemistry*; D011188:Potassium / Q000378:metabolism; D012433:Ryanodine / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel; D012964:Sodium / Q000378:metabolism
D015220:Calcium Channels; D002148:Calmodulin-Binding Proteins; D009124:Muscle Proteins; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine; D004533:Egtazic Acid; D004492:Edetic Acid; D012964:Sodium; D011188:Potassium; D002118:Calcium