Cytoskeletal restraints of band 3 rotational mobility in human erythrocyte membranes.
The interaction of band 3 with cytoskeletal proteins was investigated in erythrocyte membranes by measuring the rotational mobility of band 3 using the method of transient dichroism. It was found that selective proteolysis of ankyrin, a protein known to link band 3 to the spectrin-actin network, had no significant effect on band 3 rotation. Incubating ghosts to 70 degrees C, at which temperature ankyrin is expected to be denatured, also had no effect. It thus appears probable that linkage of band 3 to the cytoskeleton via ankyrin does not act as a restraint on band 3 rotational motion. It is suggested that this is a consequence of flexibility in the cytoskeletal structure. In further investigations of the effect of heat treatment, a large enhancement of band 3 rotational mobility was found to result from incubation of intact cells for 1 h at 50 degrees C. This effect was not observed if ghosts were subjected to the same treatment, nor did it occur if the incubation of cells was performed at 47 degrees C. These findings, in combination with previous studies of band 3 rotational mobility, indicate that the interactions which restrain band 3 are likely to be more complex than commonly envisaged.
Clague
M J
MJ
Department of Chemistry and Biological Chemistry, University of Essex, Colchester, U.K.
Harrison
J P
JP
Cherry
R J
RJ
D001457:Anion Exchange Protein 1, Erythrocyte / Q000502:physiology*; D017487:Ankyrins; D001798:Blood Proteins / Q000378:metabolism; D002942:Circular Dichroism; D003598:Cytoskeletal Proteins / Q000502:physiology*; D004910:Erythrocyte Membrane / Q000032:analysis / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D008560:Membrane Fluidity; D008565:Membrane Proteins / Q000378:metabolism; D008958:Models, Molecular; D011489:Protein Denaturation; D012399:Rotation; D013816:Thermodynamics; D014357:Trypsin
D001457:Anion Exchange Protein 1, Erythrocyte; D017487:Ankyrins; D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D014357:Trypsin