Regulation of band 3 mobilities in erythrocyte ghost membranes by protein association and cytoskeletal meshwork.
Rotational diffusion of erythrocyte anion channel protein band 3 was measured in ghost membranes by observing time-resolved phosphorescence anisotropy decays of eosinyl-5-maleimide covalently attached to the protein. Experiments were carried out under conditions similar to those employed by Tsuji and Ohnishi (1986) for translational diffusion measurement of band 3 [(1986) Biochemistry 25, 6133-6139] to allow direct comparison of rotational and translational diffusion of band 3. Detailed analysis of diffusive properties of band 3 in ghost membranes was made on the basis of these rotational and translational diffusion data. Rotational diffusion measurements indicated that there are at least three populations of band 3 molecules with high, low, and no rotational mobilities in the time scale of 10(-4)-10(-2) s. These populations are in equilibrium, and the fractional ratios are strongly temperature dependent. At 26 degrees C, 44% of band 3 molecules are mobile (16% have an average rotational correlation time of 0.19 ms, and 28% have an average correlation time of 2.4 ms), and 56% are immobile. These results correlate well with translational diffusion data which indicated 40% mobile and 60% immobile fractions of band 3. The rotational diffusion data together with the translational diffusion data by Tsuji and Ohnishi (1986) and Golan and Veatch [(1980) Proc. Natl. Acad. Sci. U.S.A. 77, 2537-2541] suggest that immobilization of band 3 is largely caused by binding of band 3 oligomers to ankyrin, which abolishes both rotational and translational diffusion of band 3.(ABSTRACT TRUNCATED AT 250 WORDS)
Tsuji
A
A
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
Kawasaki
K
K
Ohnishi
S
S
Merkle
H
H
Kusumi
A
A
D000345:Affinity Labels; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism*; D004058:Diffusion; D004591:Electrophoresis, Polyacrylamide Gel; D004801:Eosine Yellowish-(YS) / Q000031:analogs & derivatives; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D008433:Mathematics; D008565:Membrane Proteins / Q000097:blood*; D010783:Photometry; D011700:Putrescine / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013095:Spermidine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology
D000345:Affinity Labels; D001457:Anion Exchange Protein 1, Erythrocyte; D046911:Macromolecular Substances; D008565:Membrane Proteins; D013096:Spermine; D012965:Sodium Chloride; C033531:eosin maleimide; D004801:Eosine Yellowish-(YS); D013095:Spermidine; D011700:Putrescine