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  • Title: Characterization of a ryanodine receptor in Periplaneta americana.
    Author: Schmitt M, Turberg A, Londershausen M.
    Journal: J Recept Signal Transduct Res; 1997; 17(1-3):185-97. PubMed ID: 9029490.
    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.
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