一种快速大量纯化骨骼肌ryanodine受体的方法

Ryanodine受体（RyR1）在骨骼肌细胞的兴奋-收缩偶联过程中扮演重要角色，是肌质网快速释放Ca²⁺的通道。RyR1的结构研究不同于其功能研究，需要大量的且纯度很高的蛋白。通过运用肝素（Heparin，HP）层析与羟基磷灰石（Hydroxylapatite，HA）层析，不仅在一天的时间内就可以纯化获得毫克级的RyR1，而且RyR1的纯度达到95%以上。在透射电子显微镜下观察到RyR1是一个正方形的结构，边长大约26 nm，形态类似儿童玩具风车。这个方法获得RyR1速度快、纯度高、结构完整，为进行RyR1结构研究奠定了基础。

A Fast Method for the Purification of the Skeletal Muscle Ryanodine Receptor in Large Scale

Ryanodine receptor 1(RyR1) plays a vital role in excitation-contraction coupling in skeletal muscle. It is the fast Ca 2 release channel of the sarcoplasmic reticulum. Large amount of purified protein is required for structural studies of RyR1. Currently, the methods in literature are all small scale and cannot fulfill the requirement for structural studies. By using fast flow heparin(HP)and hydroxyapatite(HA)chromatography, we purified the RyR1 in milligram scale in one day. SDS-PAGE showed that the purity is over 95%; electron microscopy revealed that the purified RyR1 molecule displays a square-like structure with an appearance of a pinwheel. These data demonstrate that both purify and structural integrity is achieved by this fast method.