title: Rotational diffusion of band 3 in erythrocyte membranes. 2. Binding of cytoplasmic enzymes.
abstract: Time-resolved phosphorescence anisotropy has been used to study the rotational diffusion of eosin-labeled human erythrocyte band 3 in the presence of an enzyme bound at its cytoplasmic pole. With increasing amounts of G3PD (glyceraldehyde-3-phosphate dehydrogenase) added to ghosts, the infinite time anisotropy (r infinity) increases, and at saturating concentrations, very little decay of the anisotropy r(t) occurs at all. These phenomena are reversed by elution of the enzyme with 150 mM NaCl. Prior proteolytic removal of the N-terminal 41-kDa cytoplasmic fragment of band 3 also disenables the G3PD effect. When ghosts are stripped of their residually bound G3PD, a small reduction in the fraction of immobile band 3 is observed. Finally, titration of band 3 sites with aldolase shows effects on the r(t) qualitatively similar to those observed with G3PD. On the basis of our interpretation of the heterogenous anisotropy decay of eosin-labeled band 3 [Matayoshi, E. D., & Jovin, T. M. (1991) Biochemistry (preceding paper in this issue)], we conclude that the binding of G3PD and aldolase results in the immobilization of band 3 oligomers.
authors.affiliation: Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, FRG.
authors.forename: E D
authors.initials: ED
authors.lastname: Matayoshi
authors.forename: W H
authors.initials: WH
authors.lastname: Sawyer
authors.forename: T M
authors.initials: TM
authors.lastname: Jovin
mesh_terms: D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry*; D003593:Cytoplasm / Q000201:enzymology; D004058:Diffusion; D004910:Erythrocyte Membrane / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology*; D005454:Fluorescence Polarization; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D011817:Rabbits; D012965:Sodium Chloride / Q000494:pharmacology
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D012965:Sodium Chloride; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005634:Fructose-Bisphosphate Aldolase