Service de Biophysique, C.E.N. Saclay, 91191, Gif-sur-Yvette, France.
Photosynth Res. 1987 Jan;11(2):141-51. doi: 10.1007/BF00018272.
Chloroplasts of a chlorophyll (Chl) b-less barley mutant were solubilized with digitonin and fractionated by polyacrylamide gel electrophoresis with sodium deoxycholate in the running buffer. By this procedure, in contrast to using sodium dodecylsulfate (SDS) for solubilization, a Chl a-protein analogous to the major light-harvesting Chl a-b protein complex from wildtype chloroplasts was recovered. This mutant Chl a-protein comprises about fifty percent of the total Chl a, and is very similar in carotenoid, amino acid, protein and polypeptide composition to the major wildtype antenna Chl a-b protein. The only major differences we have found is its instability in the presence of SDS and sensitivity to protease action. Even with deoxycholate, the mutant Chl a complex often dissociates during electrophoresis into two green bands. The lack of Chl b appears to affect the normal organization of Chl a and protein in such a way as to render the complex more unstable.
用胆酸钠作离子去污剂，通过聚丙烯酰胺凝胶电泳可以将缺少叶绿素 b 的大麦突变体的叶绿体进行溶解和分级分离。与用十二烷基硫酸钠（SDS）进行溶解相比，通过这个步骤可以回收类似于野生型叶绿体主要光捕获叶绿素 a-b 蛋白复合物的叶绿素 a 蛋白。这种突变叶绿素 a 蛋白约占总叶绿素 a 的 50%，其类胡萝卜素、氨基酸、蛋白质和多肽组成与主要的野生型天线叶绿素 a-b 蛋白非常相似。我们发现的唯一主要差异是它在 SDS 存在下的不稳定性和对蛋白酶作用的敏感性。即使有胆酸钠，突变的叶绿素 a 复合物在电泳过程中也常常分解成两条绿色带。缺少叶绿素 b 似乎以一种使复合物更不稳定的方式影响叶绿素 a 和蛋白质的正常组织。
