A rapid and efficient purification method for recombinant annexin V for biophysical studies

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Abstract
Annexin V binds in a calcium‐dependent manner to acidic phospholipids and exhibits ion channel activity in vitro. We are investigating mutants of annexin V by single channel measurements, X‐ray crystallography and electron microscopy in order to understand the structure‐function relationships of the ion channel activity. We describe here a method to obtain very pure reeombinant annexin V required for such studies. The initial step is the mild opening of the bacterial cells by an osmotic shock. In the purification procedure, use is made of the reversible calcium‐mediated binding of annexin V to liposomes. In the last purification step the protein is subjected to ion‐exchange chromatography and elutes as a single peak free of any detectable contaminants.