Department of Biology, Technion-Israel Institute of Technology, Haifa 32 000, Israel.
Plant Physiol. 1985 Jul;78(3):586-90. doi: 10.1104/pp.78.3.586.
Ribulose bisphosphate carboxylase (Rubisco) has been found in Vicia faba L. guard cell chloroplasts by two immunological methods, using antibodies raised against highly purified subunits of ribulose bisphosphate carboxylase. Indirect cytoimmunofluorescence revealed binding of antibodies against both the small and the large subunits of ribulose bisphosphate carboxylase. Binding was observed only after partial digestion of guard cell walls by 4% Cellulysin to facilitate antibody penetration. After electrophoresis of a homogenate of guard cell protoplasts, the presence of both subunits was also revealed by immunolabeling technique. Positive response required the inhibition of proteolysis which appeared to be active upon homogenization.
利用针对核酮糖二磷酸羧化酶高度纯化亚基的抗体,通过两种免疫学方法,在蚕豆保卫细胞叶绿体中发现了核酮糖二磷酸羧化酶(Rubisco)。间接细胞免疫荧光显示,针对核酮糖二磷酸羧化酶的小亚基和大亚基的抗体都有结合。只有在用 4%纤维素酶部分消化细胞壁以促进抗体渗透后,才观察到结合。在保卫细胞原生质体匀浆电泳后,免疫标记技术也显示了两个亚基的存在。阳性反应需要抑制似乎在匀浆时活跃的蛋白水解。