Characterization of calcium binding to brain spectrin

Wallis, C. J.; Wenegieme, Elizabeth F.; Babitch, Joseph A.

Publication

Characterization of calcium binding to brain spectrin

Wallis, C. J.; Wenegieme, Elizabeth F.; Babitch, Joseph A.

Publisher

American Society for Biochemistry and Molecular Biology Inc.

Date

1992

Abstract

Brain spectrin alpha and beta chains bind 45Ca2+, as shown by the calcium overlay method. Flow dialysis measurements revealed eight high affinity binding sites/tetramer that comprise two binding components (determined by nonlinear regression analysis). The first component has one or two sites (kd = 2-30 x 10(-8) M), depending on the ionic strength of the binding buffer, with the remaining high affinity sites in the second component (kd = 1-3 x 10(-6) M). In addition, there is a variable, low affinity binding component (n = 100-400, kd = 1-2 x 10(-4) M). Magnesium inhibits calcium binding to the low affinity sites with a K1 = 1.21 mM. Proteolytic fragments from trypsin or chymotrypsin digests of brain spectrin bind 45Ca2+ if they include alpha domain IV, alpha domain III, or the amino-terminal half of the beta chain (but more than 25 kDa from the amino-terminal). These data suggest that calcium ions bind with high affinity to the putative EF-hands in alpha domain IV and to one site in the amino-terminal half of the beta chain that is associated with alpha domain IV in the native dimer. The localization is consistent with a direct calcium modulation of the spectrin-actin-protein 4.1 interaction. In addition, there appears to be one high affinity site near the hypersensitive region of alpha brain spectrin. All four proposed binding sites occur near probable calmodulin-binding or calcium-dependent protease cleavage sites.

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Department

Chemistry and Biochemistry

URI

https://repository.tcu.edu/handle/116099117/56606

DOI

https://doi.org/10.1016/s0021-9258(18)42839-6

Characterization of calcium binding to brain spectrin

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