Use of monoclonal antibodies to study acetylcholine receptors from electric organs, muscle, and brain and the autoimmune response to receptor in myasthenia gravis.
Use of monoclonal antibodies to study acetylcholine receptors from electric organs, muscle, and brain and the autoimmune response to receptor in myasthenia gravis.
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使用单克隆抗体研究来自电器官、肌肉和大脑的乙酰胆碱受体以及重症肌无力受体的自身免疫反应。
DOI：
10.1101/sqb.1983.048.01.012
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Montal,M
中科院分区：
文献类型：
--
作者：
Lindstrom,J;Tzartos,S;Gullick,W;Hochschwender,S;Swanson,L;Sargent,P;Jacob,M;Montal,M
单克隆抗体在乙酰胆碱受体 (AChR) 和 AChR 自身免疫反应的研究中有许多应用。它们很有用，因为它们可以为受体分子提供各种高度特异性的探针，这些探针适合多种类型的实验操作，并且因为它们可以用作模型自身抗体。我们在这里描述的基本方法和技术在神经生物学家感兴趣的许多其他受体和分子的研究中应该被证明是有价值的。我们已经生成了一个包含 160 多种针对天然 AChR 的单克隆抗体的文库，这些抗体是从加州鱼雷 (Tzartos 和 Lindstrom 1980) 和电电鱼 (Tzartos et al. 1981) 的电器官以及牛的肌肉 (Tzartos 和 Lindstrom 1981; S. Tzartos 等人，未发表) 和人类 (Tzartos 等人，1981) 的肌肉中纯化出来的。 1983） 以及来自电器官（Tzartos 和 Lindstrom 1980；S. Tzartos 等人，未发表）和肌肉（S. Hochschwender 等人，未发表）的受体变性亚基。为了获得我们需要的所有单克隆抗体，我们正在继续生成和表征其他精心挑选的抗受体单克隆抗体。这种生成并仔细表征针对单一类型受体的大型单克隆抗体库的密集方法非常费力，并且与神经生物学中经常使用的用原始组织进行免疫并使用出现的任何有趣的细胞特异性单克隆抗体的方法完全不同。然而，这种策略允许进行许多类型的实验，否则这些实验是不可能的，或者结果不会那么明确。我们使用单克隆抗体研究电器官组织和肌肉中的 AChR 至少有五类应用：(1) 识别受体亚基并定位受体亚基内的亚基，(2) 测试它们结合的亚基结构在受体功能中的作用，(3) 比较来自不同组织和物种的受体的结构，(4) 定位受体四级结构内的亚基。 受体分子，(5) 研究受体亚基的合成、构象成熟和组装。本文回顾了前三个应用中涉及的一些进展和当前方法，这些应用使用单克隆抗体来研究电器官和 mus-89 cle 受体的结构和功能。提供了一些关于抗原决定簇的跨膜方向以及结合的单克隆抗体对受体功能的影响的数据。针对来自 T. californica 电器官的受体的亚基特异性单克隆抗体正在 Stroud 等人（本卷）描述的受体结构的优雅高分辨率电子显微镜研究中使用。 Anderson 等人（1983 年和本卷）描述了使用单克隆抗体来确定体外合成的鱼雷受体亚基的早期生物合成形式中抗原决定簇的跨膜方向。 Merlie 等人（1982 年、1983 年和本卷；Sebbane 等人 1983 年；JP Merlie 和 J. Lindstrom，准备中。1983 年）描述了使用单克隆抗体来研究组织培养物中生长的肌肉细胞中受体亚基的合成、构象成熟和组装。亚基特异性单克隆抗体是识别新合成的受体亚基的关键工具，并且它们提供了检测其构象成熟和组装的方法。单克隆抗体在重症肌无力对 AChR 的自身免疫反应研究中具有三种主要应用……
Monoclonal antibodies have many applications in the study of acetylcholine receptors (AChRs) and the autoimmune response to AChRs. They are useful because they can provide a wide range of highly specific probes for the receptor molecule that are amenable to many types of experimental manipulation, and because they can be used as model autoantibodies. The basic approaches and techniques that we describe here should prove valuable in the study of many other receptors and molecules of interest to neurobiologists. We have generated a library of more than 160 monoclonal antibodies to native AChRs purified from the electric organs of Torpedo californica (Tzartos and Lindstrom 1980) and Electrophorus electricus (Tzartos et al. 1981) and the muscles of cattle (Tzartos and Lindstrom 1981; S. Tzartos et al., unpubl.) and humans (Tzartos et al. 1983) and to the denatured subunits of receptors from both electric organs (Tzartos and Lindstrom 1980; S. Tzartos et al., unpubl.) and muscle (S. Hochschwender et al., unpubl.). To obtain all of the monoclonal antibodies we require, we are continuing to generate and characterize other carefully selected antireceptor monoclonal antibodies. This intensive approach of generating and carefully characterizing a large library of monoclonal antibodies to a single type of receptor is very laborious, and quite different from the approach frequently used in neurobiology of immunizing with crude tissue and using any interesting cell-specific monoclonal antibodies that arise. However, this strategy permits many types of experiments that would otherwise not be possible or where the resuits would not be so unambiguous. There are at least five classes of applications in which we are using monoclonal antibodies for studies of AChRs from electric organ tissue and muscle:(1) to identify receptor subunits and localize substructures within receptor subunits,(2) to test the role of the substructures to which they bind in receptor function,(3) to compare the structures of receptors from different tissues and species,(4) to localize subunits within the quaternary structure of the receptor molecule, and (5) to study the synthesis, conformational maturation, and assembly of receptor subunits. This paper reviews some of the progress and current approaches involved in the first three applications listed that use monoclonal antibodies to study the structure and function of receptors from electric organ and mus-89 cle. Some data are presented on the transmembrane orientation of antigenic determinants and the effects of bound monoclonal antibodies on receptor function. Subunit-specific monoclonal antibodies to receptor from T. californica electric organ are being employed in the elegant high-resolution electron microscopy studies of receptor structure described by Stroud et al.(this volume). Anderson et al.(1983 and this volume) describe the use of monoclonal antibodies to determine the transmembrane orientation of antigenic determinants in early biosynthetic forms of subunits of receptor from Torpedo synthesized in vitro. Merlie et al.(1982, 1983, and this volume; Sebbane et al. 1983; JP Merlie and J. Lindstrom, in prep. 1983) describe the use of monoclonal antibodies to study the synthesis, conformational maturation, and assembly of subunits of receptor in muscle cells grown in tissue culture. Subunit-specific monoclonal antibodies are the critical tool that makes it possible to recognize newly synthesized receptor subunits, and they provide methods for detecting their conformational maturation and assembly. There are three main applications of monoclonal antibodies in the study of the autoimmune response to AChRs in myasthenia gravis …