Synergistic Effects of Copper Sites on Apparent Stability of Multicopper Oxidase, Fet3p
AbstractSaccharomyces cerevisiae Fet3p is a multicopper oxidase that contains three cupredoxin-like domains and four copper ions located in three distinct metal sites (T1 in domain 3; T2 and the binuclear T3 at the interface between domains 1 and 3). To probe the role of the copper sites in Fet3p thermodynamic stability, we performed urea-induced unfolding experiments with holo-, apo- and three partially-metallated (T1, T2 and T1/T2 sites depleted of copper) forms of Fet3p. Using a combination of spectroscopic probes (circular dichroism, fluorescence intensity and maximum, 8-anilinonaphthalene-1-sulfonic acid (ANS) emission, oxidase activity and blue color), we reveal that all forms of Fet3p unfold in a four-state reaction with two partially-folded intermediates. Using phase diagrams, it emerged that Fet3p with all copper sites filled had a significantly higher stability as compared to the combined contributions of the individual copper sites. Hence, there is long-range inter-domain communication between distal copper sites that contribute to overall Fet3p stability. View Full-Text
- Supplementary File 1:
Supplementary (PDF, 277 KB)
Scifeed alert for new publicationsNever miss any articles matching your research from any publisher
- Get alerts for new papers matching your research
- Find out the new papers from selected authors
- Updated daily for 49'000+ journals and 6000+ publishers
- Define your Scifeed now
Sedlák, E.; Žoldák, G.; Wittung-Stafshede, P. Synergistic Effects of Copper Sites on Apparent Stability of Multicopper Oxidase, Fet3p. Int. J. Mol. Sci. 2018, 19, 269.
Sedlák E, Žoldák G, Wittung-Stafshede P. Synergistic Effects of Copper Sites on Apparent Stability of Multicopper Oxidase, Fet3p. International Journal of Molecular Sciences. 2018; 19(1):269.Chicago/Turabian Style
Sedlák, Erik; Žoldák, Gabriel; Wittung-Stafshede, Pernilla. 2018. "Synergistic Effects of Copper Sites on Apparent Stability of Multicopper Oxidase, Fet3p." Int. J. Mol. Sci. 19, no. 1: 269.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.