An immunologic approach to the conformational equilibria of polypeptides.
An immunologic approach to the conformational equilibria of polypeptides.
复制标题
多肽构象平衡的免疫学方法。
DOI：
10.1073/pnas.69.12.3790
复制
发表时间：
1972
影响因子：
11.1
通讯作者：
C. Anfinsen
中科院分区：
文献类型：
--
作者：
D. Sachs;A. Schechter;A. Eastlake;C. Anfinsen
用核酸酶的多肽片段免疫和用完整的核酸酶免疫，然后在带有相应片段的免疫吸收柱上分离抗血清，制备了与葡萄球菌核酸酶分子不同区域有反应的抗体。通过定量沉淀测定和酶抑制研究，比较这些抗体制剂与核酸酶的相互作用，发现由于免疫抗原的构象存在显著差异。提出了一种解释模型，其中从抗核酸酶血清中分离出的抗体仅与核酸酶多肽片段的“天然格式决定因素”有效反应。假设这些决定因子是在多肽片段中通过多肽链的自发和可逆折叠产生的。该模型允许实验确定所提出的构象平衡的参数K（conf）。
Antibodies reactive with distinct regions of the staphylococcal nuclease molecule were prepared both by immunization with polypeptide fragments of nuclease and by immunization with intact nuclease followed by fractionation of the antiserum on immuno-absorbent columns bearing the corresponding fragments. Comparisons of the interactions of these antibody preparations with nuclease, by quantitative precipitin assays and enzyme inhibition studies, showed marked differences attributable to the conformation of the immunizing antigens. An interpretive model is proposed in which antibodies fractionated from anti-nuclease serum react effectively only with the "native format determinants" of polypeptide fragments of nuclease. It is postulated that such determinants are generated in polypeptide fragments by spontaneous and reversible folding of the polypeptide chain. The model permits experimental determination of parameters, K(conf), for the proposed conformational equilibria.