Next Article in Journal
Cytokine Disturbances in Coronary Artery Ectasia Do Not Support Atherosclerosis Pathogenesis
Next Article in Special Issue
Unveiling a Selective Mechanism for the Inhibition of α-Synuclein Aggregation by β-Synuclein
Previous Article in Journal
Caffeic Acid and Metformin Inhibit Invasive Phenotype Induced by TGF-β1 in C-4I and HTB-35/SiHa Human Cervical Squamous Carcinoma Cells by Acting on Different Molecular Targets
Previous Article in Special Issue
Modulation of Protein Quality Control and Proteasome to Autophagy Switch in Immortalized Myoblasts from Duchenne Muscular Dystrophy Patients
Article

Synergistic Effects of Copper Sites on Apparent Stability of Multicopper Oxidase, Fet3p

1
Center for Interdisciplinary Biosciences, Technology and Innovation Park P.J. Šafárik University, Jesenna 5, 041 54 Košice, Slovakia
2
Department of Biology and Biological Engineering, Division of Chemical Biology, Chalmers University of Technology, 412 96 Gothenburg, Sweden
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2018, 19(1), 269; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms19010269
Received: 28 November 2017 / Revised: 19 December 2017 / Accepted: 12 January 2018 / Published: 16 January 2018
(This article belongs to the Collection Protein Folding)
Saccharomyces 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
Keywords: multicopper oxidases MCO; cupredoxin-like domain; phase diagram method; multidomain protein stability; Fet3p multicopper oxidases MCO; cupredoxin-like domain; phase diagram method; multidomain protein stability; Fet3p
Show Figures

Figure 1

MDPI and ACS Style

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. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms19010269

AMA Style

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. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms19010269

Chicago/Turabian Style

Sedlák, Erik, Gabriel Žoldák, and Pernilla Wittung-Stafshede. 2018. "Synergistic Effects of Copper Sites on Apparent Stability of Multicopper Oxidase, Fet3p" International Journal of Molecular Sciences 19, no. 1: 269. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms19010269

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop