A RAPID LECTIN RECEPTOR-BINDING ASSAY - COMPARATIVE-EVALUATION OF SEA-URCHIN EMBRYO CELL-SURFACE LECTIN RECEPTORS

被引:20
|
作者
LATHAM, VH
DUCUT, JL
ROSTAMIANI, K
CHUN, HH
LOPEZ, ME
HERRERA, S
OPPENHEIMER, SB
机构
[1] CALIF STATE UNIV NORTHRIDGE,CTR CANC & DEV BIOL,NORTHRIDGE,CA 91330
[2] CALIF STATE UNIV NORTHRIDGE,DEPT BIOL,NORTHRIDGE,CA 91330
基金
美国国家卫生研究院;
关键词
LECTIN RECEPTOR BINDING ASSAY; SEA URCHIN EMBRYO;
D O I
10.1016/S0065-1281(11)80209-6
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Lectin receptor binding assays, such as those that utilize fluorescence, radioactivity or electron microscopy are not designed for rapidly screening hundreds of cell types for the presence or absence of specific lectin receptors. An assay is described here that is designated for this purpose. It utilizes lectins derivatized to agarose beads and can be used to screen many cell types in min. This assay was used to examine lectin receptors on the surfaces of 1-8 cell stage Strongylocentrotus purpuratus sea urchin embryos. The same cells were also assayed using standard fluorescence and agglutinability procedures to ascertain the type of information obtained by the new assay and how it correlates with results from the standard methods. The bead results correlated well with results using fluorescent lectin. Only wheat germ agglutinin bound very strongly in both bead and fluorescence assays, while concanavalin A, Dolichos biflorus, Lens culinaris and Tetragonolobus purpureas did not bind or bound weakly using both methods. Results using a third method, lectin mediated cell agglutination, did not correlate with the bead or fluorescence assays. Lectin receptors were also examined on embryos prepared by two different methods of preventing formation of fertilization membranes, so that coat-free cell surfaces could be studied, the standard dithiothreitol method and a new method using alpha-amylase. Lectin receptors on the cell surfaces of embryos prepared by both methods were nearly identical. The possible functions of WGA receptors, the most prevalent lectin receptors of those studied, that were uniformly present throughout early development of this sea urchin species, are considered. The benefits and drawbacks of the agarose bead lectin receptor binding assay are also discussed.
引用
收藏
页码:89 / 97
页数:9
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