Metallation state of human manganese superoxide dismutase expressed in Saccharomyces cerevisiae.

Archives of Biochemistry and Biophysics
Mei M Whittaker, James W Whittaker

Abstract

Human manganese superoxide dismutase (Sod2p) has been expressed in yeast and the protein purified from isolated yeast mitochondria, yielding both the metallated protein and the less stable apoprotein in a single chromatographic step. At 30 °C growth temperature, more than half of the purified enzyme is apoprotein that can be fully activated following reconstitution, while the remainder contains a mixture of manganese and iron. In contrast, only fully metallated enzyme was isolated from a similarly constructed yeast strain expressing the homologous yeast manganese superoxide dismutase. Both the manganese content and superoxide dismutase activity of the recombinant human enzyme increased with increasing growth temperatures. The dependence of in vivo metallation state on growth temperature resembles the in vitro thermal activation behavior of human manganese superoxide dismutase observed in previous studies. Partially metallated human superoxide dismutase is fully active in protecting yeast against superoxide stress produced by addition of paraquat to the growth medium. However, a splice variant of human manganese superoxide dismutase (isoform B) is expressed as insoluble protein in both Escherichia coli and yeast mitochondria and...Continue Reading

References

Jun 19, 1998·Biochemistry·M M WhittakerJ W Whittaker
Jun 22, 2006·Journal of Cell Science·Anabella Srebrow, Alberto R Kornblihtt
Jul 11, 2006·Biochimica Et Biophysica Acta·Valeria Cizewski CulottaThomas V O'Halloran
Oct 10, 2008·Biochemistry·Mei M Whittaker, James W Whittaker
May 1, 2009·Cell Biochemistry and Function·Daniel Hernandez-Saavedra, Joe M McCord

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Citations

Nov 5, 2013·Protein Expression and Purification·Mei M Whittaker, James W Whittaker
Apr 20, 2013·Metallomics : Integrated Biometal Science·Jinkyu ParkPaul A Lindahl
Feb 5, 2013·The Journal of Biological Chemistry·J Dafhne AguirreValeria C Culotta
Feb 1, 2015·Archives of Biochemistry and Biophysics·Mei M WhittakerJames W Whittaker
Apr 8, 2021·Nature Communications·Jahaun AzadmaneshGloria E O Borgstahl
Apr 2, 2014·Chemical Reviews·Yuewei ShengJoan Selverstone Valentine

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