title: Characterisation of zinc-binding domains of peroxisomal RING finger proteins using size exclusion chromatography/inductively coupled plasma-mass spectrometry.
abstract: We determined the zinc binding stoichiometry of peroxisomal RING finger proteins by measuring sulfur/metal ratios using inductively coupled plasma-mass spectrometry coupled to size exclusion chromatography, a strategy that provides a fast and quantitative overview on the binding of metals in proteins. As a quality control, liquid chromatography-electrospray ionisation-time of flight-mass spectrometry was used to measure the molar masses of the intact proteins. The RING fingers of Pex2p, Pex10p, and Pex12p showed a stoichiometry of 2.0, 2.1, and 1.2 mol zinc/mol protein, respectively. Thus, Pex2p and Pex10p possess a typical RING domain with two coordinated zinc atoms, whereas that of Pex12p coordinates only a single zinc atom.
authors.affiliation: Abteilung für Analytische Chemie, Universität für Bodenkultur Wien, A-1190, Wien, Austria.
authors.forename: Gunda
authors.initials: G
authors.lastname: Koellensperger
authors.forename: Simon
authors.initials: S
authors.lastname: Daubert
authors.forename: Ralf
authors.initials: R
authors.lastname: Erdmann
authors.forename: Stephan
authors.initials: S
authors.lastname: Hann
authors.forename: Hanspeter
authors.initials: H
authors.lastname: Rottensteiner
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002850:Chromatography, Gel / Q000379:methods*; D017931:DNA Primers; D008969:Molecular Sequence Data; D020675:Peroxisomes / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D015032:Zinc / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D015032:Zinc