Wheat germ agglutinin stabilization of erythrocyte shape: role of bilayer balance and the membrane skeleton.
The effects of wheat germ agglutinin (WGA), Limulus lectin, and concanavalin A on cell shape changes were examined in human erythrocytes. These agents inhibited echinocytosis in cells having elevated cytosolic Ca2+ or incorporated foreign phosphatidylcholine, but had no effect on cell stomatocytosis in response to incorporated phosphatidylserine. The role of the membrane skeleton in this selective membrane fixation was examined. WGA inhibited echinocytosis in cells previously depleted of polyphosphoinositides to reduce membrane skeleton binding to transmembrane proteins, treated with phorbol ester to enhance protein 4.1 phosphorylation, heat-treated to denature spectrin, alkylated with p-chloromercuribenzoate to dissociate glycophorin from the membrane skeleton, or subjected to elevated cell 2,3-diphosphoglycerate to alter organization of the spectrin-actin-protein 4.1 complex. Limulus lectin and increased concentrations of WGA also stabilized discoid shape in pronase-digested cells containing no detectable intact glycophorin. In contrast, cell digestion with sialidase abolished the shape-stabilizing effect of WGA. The results suggest that the membrane skeleton is not involved in WGA shape stabilization. Rather, they suggest that glycoproteins and glycolipids interact with the lectin to stabilize cell surface molecular associations, forming a superficial calyx that inhibits outward, but not inward, membrane bending.
Lin
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Department of Chemistry, Stanford University, CA 94305.
Huestis
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WH
D000328:Adult; D060829:Arthropod Proteins; D001798:Blood Proteins / Q000187:drug effects / Q000502:physiology*; D048429:Cell Size / Q000187:drug effects / Q000502:physiology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003208:Concanavalin A / Q000494:pharmacology; D003598:Cytoskeletal Proteins; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000502:physiology; D004912:Erythrocytes / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D005404:Fixatives; D005976:Glutaral; D006021:Glycophorins / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D037102:Lectins / Q000494:pharmacology; D008565:Membrane Proteins / Q000187:drug effects / Q000502:physiology; D009479:Neuropeptides; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates / Q000502:physiology; D010766:Phosphorylation; D011489:Protein Denaturation; D013049:Spectrin / Q000502:physiology; D014909:Wheat Germ Agglutinins / Q000494:pharmacology*; D020245:p-Chloromercuribenzoic Acid
D060829:Arthropod Proteins; D001798:Blood Proteins; D002729:Chloromercuribenzoates; D003598:Cytoskeletal Proteins; D005404:Fixatives; D006021:Glycophorins; D037102:Lectins; D008565:Membrane Proteins; D009479:Neuropeptides; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D014909:Wheat Germ Agglutinins; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; D003208:Concanavalin A; D013049:Spectrin; D020245:p-Chloromercuribenzoic Acid; D005976:Glutaral