Immunochemical and crystallographic studies of antibody D1.3 in its free, antigen-liganded, and idiotope-bound states.
Immunochemical and crystallographic studies of antibody D1.3 in its free, antigen-liganded, and idiotope-bound states.
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对游离、抗原配体和独特位结合状态的抗体 D1.3 进行免疫化学和晶体学研究。
DOI：
10.1101/sqb.1989.054.01.030
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Tello,D
中科院分区：
文献类型：
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作者：
Bentley,GA;Bhat,TN;Boulot,G;Fischmann,T;Navaza,J;Poljak,RJ;Riottot,MM;Tello,D
来自人免疫球蛋白和鼠单抗(MAb)的几个Fab片段的三维结构已通过X射线结晶学确定(有关综述，请参阅Amzel和Poljak 1979；Davies和Metzger 1983；阿尔扎里等人等人)。(1988年)。进一步分析抗体抗原结合部位的立体结构是通过对两个Fab与半抗原磷胆碱的络合物的X射线衍射研究来实现的(西格尔等人。1974年)和维生素K1OH(Amzel等人1974年)。尽管这些研究对我们目前对抗体结构和特异性的理解做出了很大贡献，但与抗原-抗体反应有关的几个问题仍然没有答案，直到最近用X射线结晶学技术对Fab-抗原复合体的结构进行研究(Amit等人)。1986年；科尔曼等人。1987年；治安官等人。1987)。这些研究集中在定义抗原决定簇(或表位)的地形或不连续性质、抗原-抗体界面的区域以及它们相互作用的化学性质(参见Colman 1988；Davies等人)。(1988年)。然而，一个尚未解决的问题是，抗体分子在与抗原结合时发生构象变化的可能性。这在很大程度上是由于在结晶未连接的Fabs以提供其游离型和抗原结合型之间必要的结构比较方面缺乏成功。一种模型抗原，蛋清溶菌酶(HEL)已在我们的实验室中用于获得在BALB/c小鼠的二次反应过程中产生的单抗(Harper等人)。1987)。这些抗体已被系统地用于探索Fab-HEL络合物的可能结晶条件。现在已经获得了几种结晶的抗原-抗体复合体(Mariuzza等人)。1983年；Fischmann等人。1988年)，包括一种具有交叉反应抗原的山鸡蛋白溶菌酶(Guillon等人)。1987)。成功地对其中之一的FabD1进行了X射线结晶学分析。3-HEL复合体(Amit et ai.1985,1986)，促使我们加紧努力使未连接的FabD1结晶。3确定与抗原结合的抗体是否发生显著构象变化。在本文中，我们报道了FabD_1的结晶。3和目前的进展情况
The three-dimensional structure of several Fab fragments from human immunoglobulins and murine monoclonal antibodies (MAbs) have been determined by X-ray crystallography (for review, see Amzel and Poljak 1979; Davies and Metzger 1983; Alzari et al. 1988). Further analyses of the steric structure of the antigen-combining site of antibodies were made possible by the X-ray diffraction studies of two Fab complexes with the haptens phosphorylcholine (Segal et al. 1974) and vitamin K1OH (Amzel et al. 1974). Although these studies have contributed much to our current understanding of antibody structure and specificity, several questions relating to antigen-antibody reactions remained unanswered until the recent structural studies of Fab-antigen complexes by X-ray crystallographic techniques (Amit et al. 1986; Colman et al. 1987; Sheriff et al. 1987). These studies converged in defining the topographical or discontinuous nature of antigenic determinants (or epitopes), the area of the antigen-antibody interface, and the chemical nature of their interactions (for review, see Colman 1988; Davies et al. 1988). A question that has remained unresolved, however, concerns the possibility of conformational changes in the antibody molecule upon binding to antigen. This has been largely due to the lack of success in crystallizing unliganded Fabs to provide the necessary structural comparison between their free and antigen-bound forms.A model antigen, hen egg white lysozyme (HEL), has been used in our laboratory to obtain monoclonal antibodies produced during the course of a secondary response in BALB/c mice (Harper et al. 1987). These antibodies have been systematically employed to explore possible crystallization conditions of Fab-HEL complexes. Several crystalline antigen-antibody complexes have now been obtained (Mariuzza et al. 1983; Fischmann et al. 1988), including one with a crossreacting antigen, pheasant egg white lysozyme (Guillon et al. 1987). The successful X-ray crystallographic analysis of one of these, the FabD1. 3-HEL complex (Amit et ai. 1985, 1986), prompted us to intensify efforts to crystallize the unliganded FabD1. 3 to establish whether significant conformational changes do occur in the antibody upon binding antigen. In this paper, we report the crystallization of FabD1. 3 and the current progress of