Improved lectin-mediated immobilization of human red blood cells in superporous agarose beads.
A new type of agarose bead, superporous agarose, was used as a gel support for immobilization of human red blood cells (RBCs) mediated by wheat germ lectin. The number of immobilized cells was similar to that obtained with commercial wheat germ lectin-agarose but the cell stability appeared to be superior. This allowed improved frontal affinity chromatographic analyses of cytochalasin B (CB)-binding to the glucose transporter GLUT1 which established a ratio of one CB-binding site per GLUT1 dimer for both plain RBCs or those treated with different poly amino acids. The measured dissociation constants, 70+/-14 nM for CB and 12+/-3 mM for glucose binding to GLUT1, are similar to those reported earlier.
Gottschalk
Ingo
I
Department of Biochemistry, Biomedical Center, Uppsala University, SE-751 23, Uppsala, Sweden. ingo.gottschalk@biokem.uu.se
Gustavsson
Per Erik
PE
Ersson
Bo
B
Lundahl
Per
P
D004912:Erythrocytes / Q000378:metabolism; D051272:Glucose Transporter Type 1; D006801:Humans; D037102:Lectins; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D012685:Sepharose
D051272:Glucose Transporter Type 1; D037102:Lectins; D009004:Monosaccharide Transport Proteins; C495677:SLC2A1 protein, human; D012685:Sepharose