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EXAFS analysis of a human Cu,Zn SOD isoform focused using non-denaturing gel electrophoresis

Sylviane Chevreux1, Pier Lorenzo Solari2, Stéphane Roudeau1, Guillaume Deves1, Isabelle Alliot3,4, Denis Testemale3,5, Jean Louis Hazemann3,5 and Richard Ortega1

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Isoelectric point isoforms of a metalloprotein, copper-zinc superoxide dismutase (CuZnSOD), separated on electrophoresis gels were analyzed using X-ray Absorption Spectroscopy. Mutations of this protein are involved in familial cases of amyotrophic lateral sclerosis. The toxicity of mutants could be relied to defects in the metallation state. Our purpose is to establish analytical protocols to study metallation state of protein isoforms such as those from CuZnSOD. We previously highlighted differences in the copper oxidation state between CuZnSOD isoforms using XANES. Here, we present the first results for EXAFS analyses performed at Cu and Zn K-edge on the majoritary expressed isoform of human CuZnSOD separated on electrophoresis gels.


PACS

87.64.kd X-ray and EXAFS

87.15.M- Spectra of biomolecules

87.14.E- Proteins

82.70.Gg Gels and sols

87.15.Tt Electrophoresis

Subjects

Soft matter, liquids and polymers

Medical physics

Biological physics

Chemical physics and physical chemistry

Dates

Issue 1 (2009)



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