RUI: The Use of Antibodies to Study the Topology of Thylakoid Protein Complexes

表面特异性的多克隆和单克隆抗体 菠菜中三种蛋白质复合物的决定簇 类囊体膜将通过亲和吸附分离 到类囊体膜的正面或反面。 这边- 将使用特异性抗体来鉴定这些蛋白质 其显示针对水环境的抗原决定簇， 而类囊体是悬浮的。 比较那些 基质暴露的蛋白质 显示管腔暴露将指示哪些蛋白质是 穿膜的 在其他实验中，单克隆抗体 将被用来证明哪些蛋白质位于 四种腔侧外源蛋白， 水氧化 这些实验将大大 增强我们对所选拓扑结构的理解 蛋白质复合物，从而允许开发 日益复杂的结构/功能模型 光合能量守恒过程。 %%% 这些实验的总体目标是增加我们的 理解横向方向和拓扑结构 光合作用能量所需的三种蛋白质复合物 偶合器. 拟议的研究将促进发展 更复杂和拓扑正确的模型， 这些蛋白质复合物，并导致更好地了解 光合作用

Polyclonal and monoclonal antibodies specific for surface determinants of three protein complexes isolated from spinach thylakoid membranes will be isolated by affinity adsorption to rightsideout or insideout thylakoid membranes. These side- specific antibodies will be used to identify those proteins which display antigenic determinants to the aqueous milieu in which the thylakoids are suspended. Comparison of those proteins with stromal exposures with those proteins displaying lumen exposures will indicate which proteins are transmembranous. In other experiments, monoclonal antibodies will be used to demonstrate which proteins lie beneath the four lumen side extrinsic proteins which are involved with water oxidation. Together, these experiments will greatly enhance our understanding of the topology of the selected protein complexes, thus allowing the development of increasingly sophisticated structure/function models of the process of photosynthetic energy conservation. %%% The overall objective of these experiments is to increase our understanding of the transverse orientations and topologies of three protein complexes required for photosynthetic energy coupling. The proposed research will enhance the development of more sophisticated and topologically correct models of these protein complexes and lead to a better understanding of photosynthesis.