Masojidek J, Droppa M, Horváth G
Institute of Plant Physiology, Biological Research Center, Szeged, Hungary.
Eur J Biochem. 1987 Dec 1;169(2):283-8. doi: 10.1111/j.1432-1033.1987.tb13609.x.
The polypeptide composition of whole thylakoids and membrane subfragments was studied by using a modified two-dimensional gel electrophoresis technique of O'Farrell [J. Biol. Chem. 250, 4007-4021 (1975)]. The modifications were lithium dodecyl sulphate solubilization instead instead of SDS, reverse isofocusing and sensitive silver staining procedure. This high-resolution technique allowed us to separate and identify about 170 polypeptides of thylakoid membranes. After separating grana and stroma thylakoids it was found that both types of lamellae contained nearly equal amounts of polypeptides, but about 70 polypeptides were different in the two preparations. In grana thylakoids, 54 polypeptides out of 95 were found to be mainly present in grana and 31 of them were only present in grana preparations. In stroma membranes, 43 polypeptides out of 99 were mainly present in stroma lamellae and 38 of these polypeptides were exclusively present in stroma lamellae. In a functional photosystem II preparation, 61 individual polypeptides could be distinguished. Most of these polypeptides were present in both grana and stroma lamellae, but 22 of them were more pronounced in grana than in stroma lamellae. 9 polypeptides of photosystem II were distinctly different in grana and stroma lamellae, and these differences may connect closely with the functional differences of photosystem II in the two types of thylakoids.
采用奥法雷尔改进的二维凝胶电泳技术[《生物化学杂志》250, 4007 - 4021 (1975)]研究了整个类囊体和膜亚片段的多肽组成。改进之处在于用十二烷基硫酸锂溶解代替SDS、反向等聚焦和灵敏的银染程序。这种高分辨率技术使我们能够分离并鉴定出类囊体膜的约170种多肽。分离基粒类囊体和基质类囊体后发现，这两种类型的片层所含多肽量几乎相等，但两种制剂中有约70种多肽不同。在基粒类囊体中，95种多肽中有54种主要存在于基粒中，其中31种仅存在于基粒制剂中。在基质膜中，99种多肽中有43种主要存在于基质片层中，其中38种仅存在于基质片层中。在功能性光系统II制剂中，可以区分出61种单个多肽。这些多肽大多数同时存在于基粒和基质片层中，但其中22种在基粒中比在基质片层中更明显。光系统II的9种多肽在基粒和基质片层中明显不同，这些差异可能与两种类囊体中光系统II的功能差异密切相关。
