Multimodal analysis of metals in copper-zinc superoxide dismutase isoforms separated on electrophoresis gels
LOBINSKI, Ryszard
Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux [IPREM]
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Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux [IPREM]
Language
en
Article de revue
This item was published in
Biochimie. 2009, vol. 91, n° 10, p. 1324--1327
Elsevier
English Abstract
It has been suggested that copper-zinc superoxide dismutase (CuZnSOD) isoforms of distinct isoelectric point (pI) could result from differences in their metallation state. Our aim was then to develop and validate analytical ...Read more >
It has been suggested that copper-zinc superoxide dismutase (CuZnSOD) isoforms of distinct isoelectric point (pI) could result from differences in their metallation state. Our aim was then to develop and validate analytical methods for the determination and understanding of metallation states in human CuZnSOD isoforms. To avoid metal losses during sample preparation steps, CuZnSOD isoforms were separated according to their pI using non-denaturing isoelectric focusing (IEF) gel electrophoresis. Metal quantification was directly performed in-gel. Cu/Zn ratios of CuZnSOD isoforms were quantified by Particle-Induced X-ray Emission (PIXE) and Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS). Cu/Zn ratios were measured close to the value of 1 as expected from the known stoichiometry of CuZnSOD with slight, but statistically significant, differences between acidic and basic isoforms. Overall, this study demonstrates that metal quantification can be performed directly on metalloproteins separated on electrophoresis gels. © 2009 Elsevier Masson SAS. All rights reserved.Read less <
English Keywords
Isoelectric focusing
Isoforms
LA-ICP-MS
PIXE
Superoxide dismutase
Origin
Hal imported