脑线粒体膜钙通道检测方法的实验研究

Experimental study of method for detecting ICa-L in mitochondrial membrane

目的建立线粒体膜钙通道的检测方法,为分离纯化钙通道蛋白提供更可靠的手段。方法采用分光光度技术,于吸光度540nm波长下测定30例沙士鼠脑线粒体膜钙通道在不同温度(30～35℃)、pH值(6.5～7.6)、蛋白质量浓度(0.2～0.6mg/ml)下的渗透性变化,以吸光度及达到平衡所用的时间反映膜钙通道变化。结果pH值7.4、0.5mg/ml线粒体蛋白含量以及25℃为线粒体膜钙通道的最佳检测方法。结论建立了鼠脑线粒体膜钙通道的最佳检测方法;可用于进一步分离纯化钙通道蛋白。

Objective To establish a method for detecting calcium channel in to provide more relioble method for the spearation and purfication of calcium chamnel protein. Methods The changes of permeability transition-the decrease of absorbance and the time to reach equilibrium（T） of calcium in mitochondria membrane for 30 rat brains were detected with the absorbance at 540 nm, by spectrophotometer technique, under different conditions （ temperature, pH and concentration of protein） ,so as to indicate the changes of calcium channel. Results The optimal condition for measuring calcium channel are pH 7.4, 0.5 mg/ml protein of mitoehondria and temperature at 25 ℃. Conclusion Method for detecting calcium channel is established, which can be used in the spearation and purification of calcium channel protein.