Analysis of human and rabbit metallothioneins by Brdicka reaction and mass spectrometry

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F14%3A43913452" target="_blank" >RIV/62156489:43210/14:43913452 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216305:26620/14:PU127036

Výsledek na webu

<a href="https://febs.onlinelibrary.wiley.com/toc/17424658/281/s1" target="_blank" >https://febs.onlinelibrary.wiley.com/toc/17424658/281/s1</a>

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Analysis of human and rabbit metallothioneins by Brdicka reaction and mass spectrometry

Popis výsledku v původním jazyce

Metallothioneins (MTs) form a large family of evolutionarily conserved low-molecular-weight proteins (~6 kDa), found in practically all life forms, in vertebrates, invertebrates, fungi and even in plants. MT plays a special role in maintaining the homeostasis of metals essential for the proper functioning of the human body, including Zn, Cu. MT is also responsible for detoxification of heavy metals such as Cd and Hg and removal of free radicals. Mammalian MTs were separated into two charge-separable isoforms, designated as MT fractions 1 and 2. The six amino acids of rabbit MT sequence show marked differences compared with the sequences of other mammalian MTs. The biological significance for this difference remains unclear. The present study demonstrates an analytical approach of employing two detection techniques: Brdicka reaction and matrix-assisted laser desorption/ionization - mass spektrometry analysis (MALDI-MS) to characterize MTs from human and rabbit liver. By MALDI-MS, rabbit and human MT were identified and additionally to monomers of MTs (~6 kDa), which are the major peaks in mass spectrum, small signals from the dimers of MTs were also observed, which are present both in human and rabbit MT. During MT analysis by Brdicka reaction, changes in signals - RS2Co ( 1.25V which represents current response of MT complex with components of Brdicka electrolyte), Cat1 ( 1.35V) and Cat2 ( 1.48V)correspondtohydrogenevolutionfromthe supportingelectrolyte catalyzed by theMT- were observed. Concentrations ofMT-0.025, 0.5and0.1 mg/mlwere analyzed.WithdecreasingMTconcentration, RS2CoandCatsignals decreasedandshifted tomorepositive potential. Signals Cat1 and Cat2 are more exposed with decreasing MT concentration. Character of the mentionedMT signals changed with differentMTconcentrations. The best signal forhumanMTwas observed in concentration 0.025 mg/ml and for rabbitMTin concentration0.05 mg/ml. Methods used in this report allow MTs identification, permit to quantify the purity and content of its isoforms, and allow studying its quantification and polymerization.

Název v anglickém jazyce

Analysis of human and rabbit metallothioneins by Brdicka reaction and mass spectrometry

Popis výsledku anglicky

Metallothioneins (MTs) form a large family of evolutionarily conserved low-molecular-weight proteins (~6 kDa), found in practically all life forms, in vertebrates, invertebrates, fungi and even in plants. MT plays a special role in maintaining the homeostasis of metals essential for the proper functioning of the human body, including Zn, Cu. MT is also responsible for detoxification of heavy metals such as Cd and Hg and removal of free radicals. Mammalian MTs were separated into two charge-separable isoforms, designated as MT fractions 1 and 2. The six amino acids of rabbit MT sequence show marked differences compared with the sequences of other mammalian MTs. The biological significance for this difference remains unclear. The present study demonstrates an analytical approach of employing two detection techniques: Brdicka reaction and matrix-assisted laser desorption/ionization - mass spektrometry analysis (MALDI-MS) to characterize MTs from human and rabbit liver. By MALDI-MS, rabbit and human MT were identified and additionally to monomers of MTs (~6 kDa), which are the major peaks in mass spectrum, small signals from the dimers of MTs were also observed, which are present both in human and rabbit MT. During MT analysis by Brdicka reaction, changes in signals - RS2Co ( 1.25V which represents current response of MT complex with components of Brdicka electrolyte), Cat1 ( 1.35V) and Cat2 ( 1.48V)correspondtohydrogenevolutionfromthe supportingelectrolyte catalyzed by theMT- were observed. Concentrations ofMT-0.025, 0.5and0.1 mg/mlwere analyzed.WithdecreasingMTconcentration, RS2CoandCatsignals decreasedandshifted tomorepositive potential. Signals Cat1 and Cat2 are more exposed with decreasing MT concentration. Character of the mentionedMT signals changed with differentMTconcentrations. The best signal forhumanMTwas observed in concentration 0.025 mg/ml and for rabbitMTin concentration0.05 mg/ml. Methods used in this report allow MTs identification, permit to quantify the purity and content of its isoforms, and allow studying its quantification and polymerization.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

<a href="/cs/project/ED1.1.00%2F02.0068" target="_blank" >ED1.1.00/02.0068: CEITEC - central european institute of technology</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2014

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů