Ca⁺⁺ dependence of the synexin and sorcin membrane association and change in sorcin hydrophobicity.

(A) Mixtures of microvesicles and nanovesicles were lysed by 0.5% saponin in the presence of Ca⁺⁺ or EDTA. The vesicle membranes were pelleted, and aliquots of the supernatants (S) and pellets (P) were analyzed by 12% polyacrylamide gel electrophoresis/silver staining (upper panel) and immunoblotting (lower panel). (B) Mixtures of microvesicles and nanovesicles were dissolved in Triton X-114 in the presence of Ca⁺⁺ or EDTA, and phase separation was performed. Aliquots of the aqueous (aq) and detergent (dt) phases were analyzed by 12% polyacrylamide gel electrophoresis/silver staining (upper panel) and immunoblotting (lower panel).