title: Molecular characterization of metal-binding polypeptide domains by electrospray ionization mass spectrometry and metal chelate affinity chromatography.
abstract: 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).
authors.affiliation: Faculty of Chemistry, University of Konstanz, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Volz
authors.forename: F U
authors.initials: FU
authors.lastname: Bosch
authors.forename: M
authors.initials: M
authors.lastname: Wunderlin
authors.forename: M
authors.initials: M
authors.lastname: Schuhmacher
authors.forename: K
authors.initials: K
authors.lastname: Melchers
authors.forename: K
authors.initials: K
authors.lastname: Bensch
authors.forename: W
authors.initials: W
authors.lastname: Steinhilber
authors.forename: K P
authors.initials: KP
authors.lastname: Schäfer
authors.forename: G
authors.initials: G
authors.lastname: Tóth
authors.forename: B
authors.initials: B
authors.lastname: Penke
authors.forename: M
authors.initials: M
authors.lastname: Przybylski
mesh_terms: 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*
chemical_list: D007106:Immune Sera; D011994:Recombinant Proteins; D003300:Copper; D009532:Nickel; D000251:Adenosine Triphosphatases