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A rapid and efficient purification method for recombinant annexin V for biophysical studies
Author(s) -
Burger A.,
Berendes R.,
Voges D.,
Huber R.,
Demange P.
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)80185-w
Subject(s) - annexin , recombinant dna , chemistry , biophysics , liposome , ion chromatography , annexin a2 , mutant , calcium , chromatography , osmotic shock , biochemistry , in vitro , biology , organic chemistry , gene
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.