Molecular characterization of metal-binding polypeptide domains by electrospray ionization mass spectrometry and metal chelate affinity chromatography

J Chromatogr A. 1998 Mar 20;800(1):29-37. doi: 10.1016/s0021-9673(97)00877-7.

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).

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / chemistry
  • Adenosine Triphosphatases / isolation & purification*
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Blotting, Western
  • Chromatography, Affinity / methods*
  • Copper / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Immune Sera / immunology
  • Molecular Sequence Data
  • Nickel / chemistry
  • Rabbits
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Immune Sera
  • Recombinant Proteins
  • Copper
  • Nickel
  • Adenosine Triphosphatases