本实验室研究发现,小麦叶片中存在着能够降解1,5-二磷酸核酮糖羧化酶/加氧酶(Rubisco)大亚基(LSU)为51KDa的相关蛋白酶,而且该蛋白酶在其分离纯化过程中与Rubisco紧密结合。本文以小麦3E158(Triticum aestivum L.cv.3E158)和扬麦158(Triticum aestivum L. cv. Yangmai158)叶片为实验材料,通过天然梯度聚丙烯酰胺凝胶电泳(GPAGE)后切胶回收得到的Rubisco,应用SDS聚丙烯酰胺凝胶电泳(SDS-PAGE)分离Rubisco蛋白亚基及蛋白酶,分别采用SDS-PAGE、加明胶的活性SDS-PAGE鉴定蛋白质和检测蛋白酶的方式,研究了该蛋白酶的生化特性和合成的细胞定位,并对与Rubisco紧密结合的蛋白酶的分离纯化进行了进一步尝试。其主要研究结果简述如下:
1、与Rubisco大亚基降解相关的蛋白酶的生化特性
小麦粗酶提取液经过非变性梯度(5%-10%)聚丙烯酰胺凝胶电泳后,通过切胶方法回收含有Rubisco的凝胶,我们得到了纯度很高的Rubisco全酶。将这种Rubisco全酶在不同的温度下进行处理2h,发现...
【英文摘要】
In our previous research, we found a kind of protease closely binding to Rubisco (Ribulose1,5-bisphosphate carboxylase/oxygenase) could degrade its large subunit (LSU) to51KDa. On the basis of former studies, using the leaf of wheat (Triticum aestivum L.cv.3E158) and Yangmai158(Triticum aestivum L., Yangmai158) as material, Rubisco holoenzyme was first obtained by gel cut after non-denaturing gradient polyacrylamind gel electrophoresis (GPAGE), and then the degradation of LSU and characters of the protease ...