Lectin receptors on the plasma membrane of soybean cells. Binding and lateral diffusion of lectins.
Lectin receptors on the plasma membrane of soybean cells. Binding and lateral diffusion of lectins.
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大豆细胞质膜上的凝集素受体。
DOI：
10.1021/bi00285a037
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发表时间：
1983
期刊：
影响因子：
2.9
通讯作者：
Schindler,M
中科院分区：
文献类型：
--
作者：
Metcalf3rd,TN;Wang,JL;Schubert,KR;Schindler,M
托马斯N.放大图片作者：John L. Wang，* Karel R. Schubert和Melvin Schindler摘要：从大豆（SB-1细胞系）根细胞悬浮培养物制备的原生质体结合大豆凝集素（SBA）、伴刀豆球蛋白A（Con A）和麦胚凝集素（WGA）。用~（125）I标记的SBA、ConA和WGA进行的结合研究表明，这些相互作用是可饱和的和特异的。荧光显微镜显示膜标记均匀。凝集素受体复合物的流动性测定荧光再分布后光漂白。SBA和Con A的扩散常数（D）分别为5 × 10 - 11和7 × 10 - 11 cm 2 ***/s。相比之下，WGA产生3 X 10 - 10 cm 2/s的扩散常数。用SBA或Con A预处理原生质体导致WGA（Z）的迁移率降低6倍（5 × 10-11 cm 2/s）。结果表明，SBA或ConA的结合可能导致大豆质膜的改变，从而限制其他受体的迁移。对质膜受体动力学的测量促进了对作为跨膜信号传导机制的重要组成部分的横向迁移作用的研究（Cherry，1979; Edidin，1981; Peters，1981）。从这些研究中，对于膜组分的运动出现了两种不同的方案：（a）作为布朗运动的结果的横向移动性（Frye & Edidin，1970; Saffman & Delbruck，1975; Schlessinger等人，1976）和（B）细胞表面上的定向流动（Taylor等，1971; Koppel等人，1982年）。基于二维连续体中扩散通量的横向迁移率已经被表征用于大量的膜蛋白（Peters，1981），并且在较小程度上用于少数种类的脂质（Peters，1981）和糖脂（Wolf等人，1977; Schindler等人，1980年b）。一般来说，这些测量是通过使用光漂白后荧光再分布的技术（FRAP; 1 Koppel，1979; Peters，1981）进行的，并且来自密歇根州立大学生物化学系，东兰辛，密歇根48824。1983年1月12日收到。这项工作得到了美国国家科学基金会的Grant PCM-8011736、美国国立卫生研究院的Grant GM-30158和密歇根农业实验站（项目编号1276 H）的支持。JLW获得了美国癌症协会FRA-221学院研究奖的支持。密歇根州农业实验站的第10849号出版物。在相同的膜中，膜蛋白的值为10-8-10-12 cm 2/s，磷脂和糖脂的值为10-8-10-9 cm 2/s。特别令人感兴趣的是，观察到一种特定的扩散蛋白质可能有一个固定的组成部分（彼得斯，1981年）。已经提出了许多理论来解释在质膜内和细胞骨架与扩散分子相互作用的背景下的这种类型的蛋白质移动性（Edelman，1976; Schindler等人，1980 b; Koppel等人，1981年）。观察到的细胞膜组分的另一种类型的运动是定向流动，最终导致淋巴细胞上的帽形成（Taylor等人，1971）和伴刀豆球蛋白A（ConA）-受体复合物在后期或末期向J7742小鼠巨噬细胞中发育的卵裂沟的运动（Koppel等，1982年）。这种类型的蛋白质流动性已被归因于膜受体的交联，通常只发生在外源添加的配体结合到受体。一个受体运动机制不需要排除其他，因为受体。
Thomas N. Metcalf, III, John L. Wang,* Karel R. Schubert, and Melvin Schindler abstract: Protoplasts prepared from suspension cultures of root cells of Glycine max(SB-1 cell line) bound soybean agglutinin (SBA), concanavalin A (Con A), and wheat germ agglutinin (WGA). Binding studies carried out with 125I-la-beled SBA, Con A, and WGA showed that these interactions were saturable and specific. Fluorescence microscopy dem-onstrated uniform membrane labeling. The mobility of the lectin-receptor complexes was measured by fluorescence re-distribution after photobleaching. The diffusion constants {D) for SBA and Con A were 5 X 10~ n and 7 X 10" 11 cm2***/s, respectively. In contrast, WGA yielded a diffusion constant of 3 X 10 “10 cm2/s. Pretreatment of the protoplasts with either SBA or Con A resulted in a 6-fold reduction in the mobility of WGA (Z)^ 5 X 10-11 cm2/s). The resultssuggest that the binding of SBA or Con A may leadto alterations of the soybean plasma membranewhich, in turn, may restrict the mobility of other receptors. e measurement of the dynamics of plasma membrane receptors has catalyzed research into the role of lateral mobility as an important component of transmembrane signaling mechanisms (Cherry, 1979; Edidin, 1981; Peters, 1981). From these investigations, two different schemes have emerged for the motion of membrane components:(a) lateral mobility as a consequence of Brownian movement (Frye & Edidin, 1970; Saffman & Delbruck, 1975; Schlessinger et al., 1976) and (b) directional flow on the cell surface (Taylor et al., 1971; Koppel et al., 1982). Lateral mobility based on diffusional fluxes in a two-dimensional continuum has been characterized for a great number of membrane proteins (Peters, 1981), and to a lesser extent for a few varieties of lipid (Peters, 1981) and glycolipid (Wolf et al., 1977; Schindler et al., 1980b). In general, these measurements have been performed by using the technique of fluorescence redistribution after photobleaching (FRAP; 1 Koppel, 1979; Peters, 1981) and have f From the Department of Biochemistry, Michigan State University, East Lansing, Michigan 48824. Received January 12, 1983. This work was supported by Grant PCM-8011736 from the National Science Foundation, by Grant GM-30158 from the National Institutes of Health, and by the Michigan Agricultural Experiment Station (Project No. 1276H). JLW was supported by Faculty Research Award FRA-221 from the American Cancer Society. Publication No. 10849 from the Michigan Agricultural Experiment Station. yielded values of from 10-8—10-12 cm2/s for membraneproteins to 10-8—10-9 cm2/s for phospholipids and glycolipids in the same membranes. Of particular interest was the observation that a particular diffusing species of protein could have an immobile component (Peters, 1981). A number of theories have been presented to explain this type of protein mobility in thecontext of intraplasmamembrane and cytoskeletal interactions with the diffusing molecule (Edelman, 1976; Schindler et al., 1980b; Koppel et al., 1981). The other type of movement for cell membranecomponents observed was a directional flow ultimately leading to cap formation on lymphoid cells (Taylor et al., 1971) and the movement of a concanavalin A (Con A)-receptor complex during late anaphase or telophase to the developing cleavage furrow in J7742 mouse macrophages (Koppel et al., 1982). This type of protein mobility has been ascribedto cross-linking of membrane receptors that normally occurs only when ex-ogenously added ligands bind to the receptors. One receptor movement mechanism need not exclude the other, since re-ceptors …