Molecular characterization of metal-binding polypeptide domains by electrospray ionization mass spectrometry and metal chelate affinity chromatography.
Metal ion-binding of synthetic peptides containing HxH and CxxC motifs was investigated by electrospray ionization mass spectrometry (ESI-MS) and metal chelate affinity chromatography. A high affinity of Ni2+ and Cu2+ to HxH containing sequences was found. Based on their natural metal ion-binding potential it was possible to include metal affinity chromatography in the purification process of two proteins without using an additional His-tag sequence: ATPase-439, a P type ATPase from Helicobacter pylori and the amyloid precursor protein (APP).
Volz
J
J
Faculty of Chemistry, University of Konstanz, Germany.
Bosch
F U
FU
Wunderlin
M
M
Schuhmacher
M
M
Melchers
K
K
Bensch
K
K
Steinhilber
W
W
Schäfer
K P
KP
Tóth
G
G
Penke
B
B
Przybylski
M
M
D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera / Q000276:immunology; D008969:Molecular Sequence Data; D009532:Nickel / Q000737:chemistry; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D007106:Immune Sera; D011994:Recombinant Proteins; D003300:Copper; D009532:Nickel; D000251:Adenosine Triphosphatases