Functional characterization of junctional terminal cisternae from mammalian fast skeletal muscle sarcoplasmic reticulum.
Functional characterization of junctional terminal cisternae from mammalian fast skeletal muscle sarcoplasmic reticulum.
复制标题
哺乳动物快速骨骼肌肌浆网连接终末池的功能特征。
DOI：
10.1021/bi00373a028
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Fleischer,S
中科院分区：
文献类型：
--
作者：
Chu,A;Volpe,P;Costello,B;Fleischer,S
肌浆网分离的材料与方法。所有SR组分均取自雌性新西兰大白兔快速骨骼肌。根据Saito et al.（1984）的描述，分离出连接末端池和来自相同梯度（R2）的轻SR分数。根据琥珀酸-细胞色素c还原酶活性估计，连接末端池的线粒体污染为2-3% （Fleischer & Fleischer, 1967）。按照先前的描述制备分离的三联体（Mitchell et al., 1983）；使用II期手术的焦磷酸盐变体（波段4）。根据Meissner（1975）的方法制备了轻、重SR囊泡，并进行了修改，在最后离心前除了常规的盐萃取步骤外，还在第一次区域离心中加入了0.6 Mkc1步骤。Ca2+转运测定。通过几种方法测量Ca2+摄取（在没有Ca2+捕获阴离子的情况下）和负载（在磷酸盐存在的情况下）。一种金属变色指示剂（Scarpa, 1979）， antipyrylazo III （Sigma Chemical）
Materials and Methods Isolation of Sarcoplasmic Reticulum. All SR fractions were prepared from fast skeletal leg muscles of female New Zealand White rabbits. Junctional terminal cisternae and a light SR fraction from the same gradient (R2) were isolated as described by Saito et al.(1984). Mitochondrial contam-ination of junctional terminal cisternae was 2-3%, as estimated by succinate-cytochrome c reductase activity (Fleischer & Fleischer, 1967). Isolated triads were prepared as previously described (Mitchell et al., 1983); the pyrophosphate variant of a stage II procedure (band 4) was used. Light and heavy SR vesicles were prepared according to Meissner (1975), with modification, in which a 0.6 Mkc1 step was included in the first zonal centrifugation in addition to the regular salt-ex-traction step before the final centrifugation. Ca2+ Transport Assays. Ca2+ uptake (in theabsence of a Ca2+-trapping anion) and loading (in the presence of phosphate) were measured by several methods. A metallochromic indicator (Scarpa, 1979), antipyrylazo III (Sigma Chemical