2,3-Diphosphoglycerate and ATP dissociate erythrocyte membrane skeletons.

Since ATP and 2,3-diphosphoglycerate cause an increase in the lateral mobility of integral membrane proteins in the erythrocyte (Schindler, M., Koppel, D., and Sheetz, M. P. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 1457-1461), we have studied their effects on the membrane skeletal complex or shell (composed of spectrin, actin, and bands 4.1 (78,000 daltons) and 4.9 (50,000 daltons)) and its interaction with the erythrocyte membrane. Both phosphate compounds dissociated the delipidated shell complex, with half-maximal dissociation at 2.5 mM 2,3-diphosphoglycerate and 8 mM ATP, whereas equivalent concentrations of EDTA did not. Concomitant with complex dissociation, spectrin was solubilized but band 4.1 and actin remained in a complexed or polymeric form. When proteins which were involved in linking spectrin to the membrane were present on the shell, higher concentrations of the phosphate compounds still dissociated the complex but less spectrin was solubilized. Treatment of erythrocyte membranes with the same phosphate compounds caused membrane vesiculation but no proteins were solubilized. We suggest that ATP and 2,3-diphosphoglycerate, at concentrations which are normally present in erythrocytes, can weaken associations in the shell but will not dissociate the complex from membrane attachment sites.