title: The Application of Curve Fitting on the Voltammograms of Various Isoforms of Metallothioneins-Metal Complexes.
abstract: The translation of metallothioneins (MTs) is one of the defense strategies by which organisms protect themselves from metal-induced toxicity. MTs belong to a family of proteins comprising MT-1, MT-2, MT-3, and MT-4 classes, with multiple isoforms within each class. The main aim of this study was to determine the behavior of MT in dependence on various externally modelled environments, using electrochemistry. In our study, the mass distribution of MTs was characterized using MALDI-TOF. After that, adsorptive transfer stripping technique with differential pulse voltammetry was selected for optimization of electrochemical detection of MTs with regard to accumulation time and pH effects. Our results show that utilization of 0.5 M NaCl, pH 6.4, as the supporting electrolyte provides a highly complicated fingerprint, showing a number of non-resolved voltammograms. Hence, we further resolved the voltammograms exhibiting the broad and overlapping signals using curve fitting. The separated signals were assigned to the electrochemical responses of several MT complexes with zinc(II), cadmium(II), and copper(II), respectively. Our results show that electrochemistry could serve as a great tool for metalloproteomic applications to determine the ratio of metal ion bonds within the target protein structure, however, it provides highly complicated signals, which require further resolution using a proper statistical method, such as curve fitting.
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic. merlos19792003@hotmail.com.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. merlos19792003@hotmail.com.
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Merlos Rodrigo
authors.affiliation: Department of Physiology, Institute of Nutrition and Food Technology, University of Granada, Avenida Del Conocimiento S/N Biomedical Research Centre, Health Campus, 18001 Granada, Spain. jrgmolinalopez@ugr.es.
authors.forename: Jorge
authors.initials: J
authors.lastname: Molina-López
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic. anuskajj@hotmail.com.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. anuskajj@hotmail.com.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Jimenez Jimenez
authors.affiliation: Department of Physiology, Institute of Nutrition and Food Technology, University of Granada, Avenida Del Conocimiento S/N Biomedical Research Centre, Health Campus, 18001 Granada, Spain. elenamp@ugr.es.
authors.forename: Elena
authors.initials: E
authors.lastname: Planells Del Pozo
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic. Pavlina.Adam@mze.cz.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. Pavlina.Adam@mze.cz.
authors.forename: Pavlina
authors.initials: P
authors.lastname: Adam
authors.affiliation: Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University and University Hospital Motol, V Uvalu 84, CZ-150 06 Prague 5, Czech Republic. Tomas.Eckschlager@fnmotol.cz.
authors.forename: Tomas
authors.initials: T
authors.lastname: Eckschlager
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic. ZitkaO@seznam.cz.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. ZitkaO@seznam.cz.
authors.forename: Ondrej
authors.initials: O
authors.lastname: Zitka
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic. oliver@centrum.cz.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. oliver@centrum.cz.
authors.forename: Lukas
authors.initials: L
authors.lastname: Richtera
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic. vojtech.adam@mendelu.cz.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czech Republic. vojtech.adam@mendelu.cz.
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
mesh_terms: D056831:Coordination Complexes / Q000737:chemistry* / Q000378:metabolism; D004563:Electrochemistry; D004573:Electrolytes; D008668:Metallothionein / Q000737:chemistry* / Q000378:metabolism; D008670:Metals / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms; D012965:Sodium Chloride / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D056831:Coordination Complexes; D004573:Electrolytes; D008670:Metals; D020033:Protein Isoforms; D012965:Sodium Chloride; D008668:Metallothionein
keywords: electrochemistry; mass spectrometry MALDI-TOF; metallomics; metallothionein; signal resolving