Insights into the role of the unusual disulfide bond in copper-zinc superoxide dismutase

Kevin Sea, Se Hui Sohn, Armando Durazo, Yuewei Sheng, Bryan F. Shaw, Xiaohang Cao, Alexander B. Taylor, Lisa J. Whitson, Stephen P. Holloway, P. John Hart, Diane E. Cabelli, Edith Butler Gralla, Joan Selverstone Valentine

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The functional and structural significance of the intrasubunit disulfide bond in copper-zinc superoxide dismutase (SOD1) was studied by characterizing mutant forms of human SOD1 (hSOD) and yeast SOD1 lacking the disulfide bond. We determined x-ray crystal structures of metal-bound and metal-deficient hC57S SOD1. C57S hSOD1 isolated from yeast contained four zinc ions per protein dimer and was structurally very similar to wild type. The addition of copper to this four-zinc protein gave properly reconstituted 2Cu,2Zn C57S hSOD, and its spectroscopic properties indicated that the coordination geometry of the copper was remarkably similar to that of holo wild type hSOD1. In contrast, the addition of copper and zinc ions to apo C57S human SOD1 failed to give proper reconstitution. Using pulse radiolysis, we determined SOD activities of yeast and human SOD1s lacking disulfide bonds and found that they were enzymatically active at ∼10% of the wild type rate. These results are contrary to earlier reports that the intrasubunit disulfide bonds in SOD1 are essential for SOD activity. Kinetic studies revealed further that the yeast mutant SOD1 had less ionic attraction for superoxide, possibly explaining the lower rates. Saccharomyces cerevisiae cells lacking the sod1 gene do not grow aerobically in the absence of lysine, but expression of C57S SOD1 increased growth to 30-50% of the growth of cells expressing wild type SOD1, supporting that C57S SOD1 retained a significant amount of activity.

Original languageEnglish (US)
Pages (from-to)2405-2418
Number of pages14
JournalJournal of Biological Chemistry
Volume290
Issue number4
DOIs
StatePublished - Jan 23 2015

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Disulfides
Yeast
Superoxide Dismutase
Zinc
Copper
Yeasts
Metals
Pulse Radiolysis
Ions
Growth
Radiolysis
Human Activities
Superoxides
Lysine
Saccharomyces cerevisiae
Proteins
Dimers
X-Rays
Genes
Crystal structure

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Sea, K., Sohn, S. H., Durazo, A., Sheng, Y., Shaw, B. F., Cao, X., ... Valentine, J. S. (2015). Insights into the role of the unusual disulfide bond in copper-zinc superoxide dismutase. Journal of Biological Chemistry, 290(4), 2405-2418. https://doi.org/10.1074/jbc.M114.588798

Insights into the role of the unusual disulfide bond in copper-zinc superoxide dismutase. / Sea, Kevin; Sohn, Se Hui; Durazo, Armando; Sheng, Yuewei; Shaw, Bryan F.; Cao, Xiaohang; Taylor, Alexander B.; Whitson, Lisa J.; Holloway, Stephen P.; Hart, P. John; Cabelli, Diane E.; Gralla, Edith Butler; Valentine, Joan Selverstone.

In: Journal of Biological Chemistry, Vol. 290, No. 4, 23.01.2015, p. 2405-2418.

Research output: Contribution to journalArticle

Sea, K, Sohn, SH, Durazo, A, Sheng, Y, Shaw, BF, Cao, X, Taylor, AB, Whitson, LJ, Holloway, SP, Hart, PJ, Cabelli, DE, Gralla, EB & Valentine, JS 2015, 'Insights into the role of the unusual disulfide bond in copper-zinc superoxide dismutase', Journal of Biological Chemistry, vol. 290, no. 4, pp. 2405-2418. https://doi.org/10.1074/jbc.M114.588798
Sea, Kevin ; Sohn, Se Hui ; Durazo, Armando ; Sheng, Yuewei ; Shaw, Bryan F. ; Cao, Xiaohang ; Taylor, Alexander B. ; Whitson, Lisa J. ; Holloway, Stephen P. ; Hart, P. John ; Cabelli, Diane E. ; Gralla, Edith Butler ; Valentine, Joan Selverstone. / Insights into the role of the unusual disulfide bond in copper-zinc superoxide dismutase. In: Journal of Biological Chemistry. 2015 ; Vol. 290, No. 4. pp. 2405-2418.
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AU - Cao, Xiaohang

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AU - Whitson, Lisa J.

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