Structure of Antibodies Binding Similar Sites on Cytochromes

结合细胞色素上相似位点的抗体的结构

• 批准号: 9019181
• 负责人: Ronald Jemmerson
• 金额: $ 26.92万
• 依托单位: University of Minnesota Saint Paul
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-02-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9019181&HistoricalAwards=false
• 关键词: Structure; Antibodies; Binding; Similar; Sites

The physicochemical parameters which guide antibody recognition of a protein antigen will be studied employing cytochrome c (cyt) as a model antigen. Of particular interest is the basis for differences in the antibody repertoires responding to variant cyts. Monoclonal antibodies (mAbs) will be elicited against several cyts that differ by as little as one amino acid in an antigenic region. The epitopes recognized will be determined from antibody binding assays employing a number of cyt variants prepared by site-directed mutagenesis or obtained from natural sources. The amino acid sequences of the heavy and light chains of the mAbs will be deduced from the nucleotide sequences of cDNA. mAbs binding similar epitopes will be compared in terms of primary structure and any differences that occur as a result of an amino acid change in an epitope will be determined. The three-dimensional structures of mAbs showing such differences will be modeled using computer graphics as will their interaction with cyt to obtain insight into the chemistry of the interaction. Some of these models will be tested by X-ray crystallography in a related study. Preliminary data indicate relatively restricted antibody gene usage with distinct V region genes used in response to the two major antigenic regions of cyt. An effect of epitope topography on complementary determining region length is also apparent. These observations will be studied further to advance our understanding of antibody recognition of native protein antigens. The results should serve as a model for protein-protein interactions, in general.

指导抗体识别的理化参数 蛋白质抗原的研究将采用细胞色素c (cyt)作为模型抗原。 特别值得注意的是 对于抗体库中的差异， 变异细胞 将诱发单克隆抗体（mAb） 针对几个相差一个氨基酸的细胞 在抗原区域。 识别的表位将是 从采用多种抗体结合测定法测定， 通过定点诱变制备或获得的Cyt变体 从天然来源。 重链的氨基酸序列 单克隆抗体的轻链将从 cDNA的核苷酸序列。结合相似表位的mAb 将在初级结构和任何 由于氨基酸变化而发生的差异， 将确定表位。 三维 将模拟显示这种差异的mAb结构 使用计算机图形学，以及它们与CYT的相互作用， 深入了解相互作用的化学过程。 一些 这些模型将通过X射线晶体学进行测试， 相关研究。 初步数据显示， 具有不同V区基因的抗体基因使用， 对cyt的两个主要抗原区域的反应。 的效果 互补决定区上的表位拓扑 长度也是显而易见的。 将对这些观察结果进行研究 进一步推进我们对抗体识别的理解 天然蛋白质抗原。 结果应作为 蛋白质相互作用的模型。