Bloodgood R A, Salomonsky N L, Reinhart F D
Department of Anatomy and Cell Biology, University of Virginia School of Medicine, Charlottesville 22908.
Exp Cell Res. 1987 Dec;173(2):572-85. doi: 10.1016/0014-4827(87)90296-5.
Two carbohydrate-binding probes (the lectin concanavalin A and the anti-carbohydrate monoclonal antibody FMG-1) have been utilized in conjunction with fluorescence-activated cell sorting to select cell lines of Chlamydomonas reinhardtii that contain defects in cell surface-exposed glycoproteins. Two very different selection strategies (sorting cells with the lowest binding for the FMG-1 monoclonal antibody or the highest binding of concanavalin A) yield a class of mutant cells that exhibit a total lack of binding of the monoclonal antibody to cell wall and plasma membrane glycoproteins along with an increased affinity for concanavalin A. Detailed characterization of one such mutant cell line, designated L-23, is provided. The subtle glycosylation defect exhibited by this cell line does not alter the ability of the affected glycoproteins to be targeted to the flagellar membrane and does not affect the expression of flagellar surface motility, a phenomenon that appears to involve the major concanavalin A-binding glycoprotein of the flagellar membrane. This approach has general applicability for dissecting the role of carbohydrate epitopes in the targeting and function of any cell surface glycoprotein for which suitable carbohydrate probes are available.
两种碳水化合物结合探针（凝集素伴刀豆球蛋白A和抗碳水化合物单克隆抗体FMG-1）已与荧光激活细胞分选技术结合使用，以筛选莱茵衣藻中在细胞表面暴露糖蛋白方面存在缺陷的细胞系。两种截然不同的筛选策略（分选与FMG-1单克隆抗体结合力最低的细胞或与伴刀豆球蛋白A结合力最高的细胞）产生了一类突变细胞，这类细胞表现出单克隆抗体与细胞壁和质膜糖蛋白完全不结合，同时对伴刀豆球蛋白A的亲和力增加。本文提供了对其中一个命名为L-23的突变细胞系的详细表征。该细胞系表现出的细微糖基化缺陷并未改变受影响糖蛋白靶向鞭毛膜的能力，也不影响鞭毛表面运动性的表达，这一现象似乎涉及鞭毛膜中主要的伴刀豆球蛋白A结合糖蛋白。对于剖析碳水化合物表位在任何有合适碳水化合物探针的细胞表面糖蛋白的靶向和功能中的作用，这种方法具有普遍适用性。
