CHEVREUX, S.; ROUDEAU, S.; FRAYSSE, A.; CARMONA, A.; DEVÈS, Guillaume; SOLARI, P.L.; MOUNICOU, Sandra; LOBINSKI, Ryszard; ORTEGA, R.

doi:10.1016/j.biochi.2009.05.016

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CHEVREUX, S.

ROUDEAU, S.

FRAYSSE, A.
Chimie Nucléaire Analytique et Bio-environnementale [CNAB]

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Biochimie. 2009, vol. 91, n° 10, p. 1324--1327

Elsevier

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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.< Réduire

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Isoelectric focusing

Isoforms

LA-ICP-MS

PIXE

Superoxide dismutase

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Multimodal analysis of metals in copper-zinc superoxide dismutase isoforms separated on electrophoresis gels

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