Effects of subunit V antibodies on the topology of the subunit and the activity of beef heart cytochrome-c oxidase.
Effects of subunit V antibodies on the topology of the subunit and the activity of beef heart cytochrome-c oxidase.
复制标题
V亚基抗体对该亚基拓扑结构和牛心细胞色素c氧化酶活性的影响。
DOI：
10.1139/o88-138
复制
发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Chan,SH
中科院分区：
文献类型：
--
作者：
Freedman,JA;Leece,B;Cooper,CE;Nicholls,P;Chan,SH
牛心脏细胞色素氧化酶亚基V上的氧化还原敏感表位先前使用兔中产生的多克隆亚基特异性抗体进行了证明。抗体仅轻微抑制电子转移，其表位的可及性取决于膜的存在和氧化酶的氧化还原状态。本文描述了额外的制备抗体提出对亚单位V。这些抗体具有更高的亚单位特异性，它们是三倍以上的抑制电子转移，它们的结合不需要一个膜。此外，它们与洗涤剂分散的氧化酶结合的氧化还原敏感性对其分离方法敏感。我们讨论的推论，可以得出一个详细的定量比较的相互作用的两种抗体制剂与抗原在不同的环境中。比较中使用的技术可用于检查氧化酶的其他干扰物对酶的特定片段的影响。
Redox-sensitive epitopes on subunit V of beef heart cytochrome-coxidase were demonstrated previously using polyclonal subunit-specific antibodies raised in rabbits. The antibodies only slightly inhibited electron transfer, and the accessibility of their epitopes depended on the presence of a membrane and on the redox state of the oxidase. The present paper describes additional preparations of antibodies raised against subunit V. These antibodies have an even higher subunit specificity, they are more than three times as inhibitory against electron transfer, and their binding does not require a membrane. Moreover, the redox-sensitive nature of their binding to detergent-dispersed oxidase is sensitive to the method of its isolation. We discuss inferences that can be drawn from a detailed quantitative comparison of the interactions of the two antibody preparations with the antigen in different environments. The techniques used in the comparison can be used to examine other perturbants of the oxidase as to their effects on specific segments of the enzyme.