High-performance liquid chromatography analysis of spectrin oligomerization

Gel filtration chromatography has been used to analyze the oligomerization of human erythrocyte spectrin. By applying an exponentially modified Gaussi an function we have been able to resolve overlapping elution peaks. From th ese peaks it was possible to calculate the equilibrium composition of each spectrin concentration and thus also the dissociation constants describing the oligomeric process. The determined dissociation constants for tetramer formation (1.3 muM) and for hexamer formation (24 muM) agree well with othe r measurements. (C) 2001 Academic Press.