Freedman J A, Leece B, Cooper C E, Nicholls P, Chan S H
Biology Department, Syracuse University, NY 13244.
Biochem Cell Biol. 1988 Nov;66(11):1210-7. doi: 10.1139/o88-138.
Redox-sensitive epitopes on subunit V of beef heart cytochrome-c oxidase 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.
先前使用在兔子体内产生的多克隆亚基特异性抗体，证实了牛心细胞色素c氧化酶亚基V上的氧化还原敏感表位。这些抗体仅轻微抑制电子传递，其表位的可及性取决于膜的存在以及氧化酶的氧化还原状态。本文描述了针对亚基V产生的抗体的其他制备方法。这些抗体具有更高的亚基特异性，它们对电子传递的抑制作用是之前的三倍多，并且它们的结合不需要膜。此外，它们与去污剂分散的氧化酶结合的氧化还原敏感性质对其分离方法敏感。我们讨论了从两种抗体制备物在不同环境中与抗原相互作用的详细定量比较中可以得出的推论。比较中使用的技术可用于检查氧化酶的其他干扰物对酶特定片段的影响。
