Glycoproteins of the CHO cell membrane: partial fractionation of the receptors for concanavalin a and wheat germ agglutinin using a lectin immunoprecipitation technique

Juliano, R.L.; Li, G.

Biochemistry 17(4): 678-683


ISSN/ISBN: 0006-2960
PMID: 341974
DOI: 10.1021/bi00597a019
Accession: 068526631

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The lectin-binding glycoproteins of the CHO [Chinese hamster ovary] cell plasma membrane were analyzed using an immunoprecipitation technique. Membranes from cells labeled via metabolic incorporation of [3H]glucosamine were solubilized in deoxycholate, the solubilized material was treated with concanavalin A, Ricinus communis agglutinins, or wheat germ agglutinin, and the lectin-glycoprotein complexes were precipitated with specific antisera directed against the lectins. The immunoprecipitates were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate and fluorescence-enhanced autoradiography. The CHO cell membrane contained 2 major glucosamine-labeled glycoprotein classes, A and B, which migrated as diffuse bands on polyacrylamide gels, and which had apparent MW of 100,000 and 130,000, respectively. Several minor labeled components were also apparent. Lectin immunoprecipitation of solubilized CHO cell membranes with wheat germ agglutinin resulted in the precipitation of material primarily of class A, while immunoprecipitation with concanavalin A produced material of class B. Thus a degree of subfractionation of the membrane glycoproteins according to lectin-binding specificity was obtained. The utility of the lectin immunoprecipitation technique is discussed in terms of analyzing the molecular associations between subclasses of membrane glycoproteins and nonglycosylated membrane macromolecules.