Possible Involvement of Glycophorin A (GPA) Interaction with Membrane Skeleton in WGA Induced Decrease in the Deformability of Erythrocyte

摘要

Erythrocyte membrane deformability is maintained mainly by fibrous spectrin molecules that interact with other membrane proteins. WGA(wheat germ agglutinin) has been shown to decrease membrane deformability(Chasis et al). However, the molecular mechanism of the WGA effect is not well understood. In this study we wanted to prove our hypothesis that WGA binds to sialic acid of GPA and then induces changes in membrane protein interactions, resulting in the deformability of erythrocyte. Human erythrocyte which had been treated with or without sialidase were incubated with various concentrations of WGA in PBS. Deformability of these cells was measured by a laser diffraction method. Simultaneously the triton shell preparation was performed and proteins were analyzed by SDS-PAGE followed by CBB, PAS staining and immunoblotting was done using monoclonal anti-GPA(alpha) (mouse IgM Isotype). Morphology was examined using dark field microscope.