Abstract
We have purified the two functionally distinct domains of gelsolin, a Ca(2+)-dependent actin binding protein, by proteolytic cleavage and characterized their size and shape in solution by dynamic light scattering. In the absence of calcium we obtained the same translational diffusion coefficient for both fragments which are of approximately equal molecular mass. The frictional ratio fo/fexp (1.33-1.39) is similar to the value as obtained for intact gelsolin (1.37) in aqueous solution (Patkowski, A., J. Seils, H. Hinssen, and T. Dorfmüller. 1990. Biopolymers. 30:427-435), indicating a similar molecular shape for the native protein as well as for the two subdomains. Upon addition of Ca2+ the translational diffusion coefficient of the carboxyl-terminal half decreased by almost 10%, while there was no change observed for the amino terminus. This result indicates that the ligand-induced conformational change as seen for intact gelsolin is probably located on the carboxyl-terminal domain of the protein. Since gelsolin has binding sites in both domains, and the isolated amino terminus binds and severs actin in a calcium-independent manner, our results suggests that the structural transition in the carboxyl-terminal part of intact gelsolin also affects the actin binding properties of the amino-terminal half.
MeSH Terms
Animals
Calcium/pharmacology
Electrophoresis, Polyacrylamide Gel
Gelsolin/chemistry,drug effects
Light
Mathematics
Molecular Weight
Muscle, Smooth
Peptide Fragments/chemistry,isolation & purification
Protein Conformation/drug effects
Scattering, Radiation
Stomach
Swine
Chemicals
Gelsolin
Peptide Fragments
Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hellweg T
Department of Chemistry, University of Bielefeld, Germany.
Hinssen H
Eimer W
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