title: LC coupled to ESI, MALDI and ICP MS - A multiple hyphenation for metalloproteomic studies.
abstract: A new multiple detection arrangement for liquid chromatography (LC) that supplements conventional electrospray ionization (ESI) mass spectrometry (MS) detection with two complementary detection techniques, matrix-assisted laser desorption/ionization (MALDI) MS and substrate-assisted laser desorption inductively coupled plasma (SALD ICP) MS has been developed. The combination of the molecular and elemental detectors in a single separation run is accomplished by utilizing a commercial MALDI target made of conductive plastic. The proposed platform provides a number of benefits in today's metalloproteomic applications, which are demonstrated by analysis of a metallothionein mixture. To maintain metallothionein complexes, separation is carried out at a neutral pH. The effluent is split; a major portion is directed to ESI MS while the remaining 1.8% fraction is deposited onto a plastic MALDI target. Dried droplets are overlaid with MALDI matrix and analysed consecutively by MALDI MS and SALD ICP MS. In the ESI MS spectra, the MT isoform complexes with metals and their stoichiometry are determined; the apoforms are revealed in the MALDI MS spectra. Quantitative determination of metallothionein isoforms is performed via determination of metals in the complexes of the individual protein isoforms using SALD ICP MS.
authors.affiliation: Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Coufalíková
authors.affiliation: Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, CEITEC MU, Masaryk University, Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Benešová
authors.affiliation: Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, CEITEC MU, Masaryk University, Brno, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Vaculovič
authors.affiliation: Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, CEITEC MU, Masaryk University, Brno, Czech Republic.
authors.forename: Viktor
authors.initials: V
authors.lastname: Kanický
authors.affiliation: Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, CEITEC MU, Masaryk University, Brno, Czech Republic. Electronic address: preisler@chemi.muni.cz.
authors.forename: Jan
authors.initials: J
authors.lastname: Preisler
mesh_terms: D002853:Chromatography, Liquid; D007834:Lasers; D008668:Metallothionein / Q000032:analysis*; D008670:Metals; D040901:Proteomics; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D008670:Metals; D008668:Metallothionein
keywords: Electrospray ionization mass spectrometry; Inductively coupled plasma mass spectrometry; Liquid chromatography; Matrix-assisted laser desorption/ionization mass spectrometry; Metal complexes; Metallothionein isoforms