Michalak M, Dupraz P, Shoshan-Barmatz V
Department of Pediatrics, University of Alberta, Edmonton, Canada.
Biochim Biophys Acta. 1988 Apr 22;939(3):587-94. doi: 10.1016/0005-2736(88)90106-x.
[3H]Ryanodine binding to skeletal muscle and cardiac sarcoplasmic reticulum (SR) vesicles was compared under experimental conditions known to inhibit or stimulate Ca2+ release. In the skeletal muscle SR, ryanodine binds to a single class of high-affinity sites (Kd of 11.3 nM). In cardiac SR vesicles, more than one class of binding sites is observed (Kd values of 3.6 and 28.1 nM). Ryanodine binding to skeletal muscle SR vesicles requires high concentrations of NaCl, whereas binding of the drug to cardiac SR is only slightly influenced by ionic strength. In the presence of 5'-adenylyl imidodiphosphate (p[NH]ppA), increased pH, and micromolar concentration of Ca2+ (which all induce Ca2+ release from SR) binding of ryanodine to SR is significantly increased in skeletal muscle, while being unchanged in cardiac muscle. Ryanodine binding to skeletal but not to cardiac muscle SR is inhibited in the presence of high Ca2+ or Mg2+ concentrations (all known to inhibit Ca2+ release from skeletal muscle SR). Ruthenium red or dicyclohexylcarbodiimide modification of cardiac and skeletal muscle SR inhibit Ca2+ release and ryanodine binding in both skeletal and cardiac membranes. These results indicate that significant differences exist in the properties of ryanodine binding to skeletal or cardiac muscle SR. Our data suggest that ryanodine binds preferably to site(s) which are accessible only when the Ca2+ release channel is in the open state.
在已知抑制或刺激Ca2+释放的实验条件下,比较了[3H]ryanodine与骨骼肌和心肌肌浆网(SR)囊泡的结合情况。在骨骼肌SR中,ryanodine与一类高亲和力位点结合(解离常数Kd为11.3 nM)。在心肌SR囊泡中,观察到不止一类结合位点(Kd值分别为3.6和28.1 nM)。ryanodine与骨骼肌SR囊泡的结合需要高浓度的NaCl,而该药物与心肌SR的结合仅受离子强度的轻微影响。在5'-腺苷酰亚胺二磷酸(p[NH]ppA)、pH升高和微摩尔浓度的Ca2+存在时(所有这些都会诱导Ca2+从SR释放),ryanodine与骨骼肌SR的结合显著增加,而在心肌中则保持不变。在高Ca2+或Mg2+浓度存在时(已知所有这些都会抑制Ca2+从骨骼肌SR释放),ryanodine与骨骼肌而非心肌SR的结合受到抑制。钌红或二环己基碳二亚胺对心肌和骨骼肌SR的修饰会抑制骨骼肌和心肌膜中的Ca2+释放和ryanodine结合。这些结果表明,ryanodine与骨骼肌或心肌SR结合的特性存在显著差异。我们的数据表明,ryanodine优先结合于仅当Ca2+释放通道处于开放状态时才可到达的位点。