In vitro thermodynamic dissection of human copper transfer from chaperone to target protein.

PloS One
Moritz S NiemiecPernilla Wittung-Stafshede

Abstract

Transient protein-protein and protein-ligand interactions are fundamental components of biological activity. To understand biological activity, not only the structures of the involved proteins are important but also the energetics of the individual steps of a reaction. Here we use in vitro biophysical methods to deduce thermodynamic parameters of copper (Cu) transfer from the human copper chaperone Atox1 to the fourth metal-binding domain of the Wilson disease protein (WD4). Atox1 and WD4 have the same fold (ferredoxin-like fold) and Cu-binding site (two surface exposed cysteine residues) and thus it is not clear what drives metal transfer from one protein to the other. Cu transfer is a two-step reaction involving a metal-dependent ternary complex in which the metal is coordinated by cysteines from both proteins (i.e., Atox1-Cu-WD4). We employ size exclusion chromatography to estimate individual equilibrium constants for the two steps. This information together with calorimetric titration data are used to reveal enthalpic and entropic contributions of each step in the transfer process. Upon combining the equilibrium constants for both steps, a metal exchange factor (from Atox1 to WD4) of 10 is calculated, governed by a negative...Continue Reading

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Citations

Dec 5, 2013·Journal of the American Chemical Society·Pritha BagchiChristoph J Fahrni
May 16, 2014·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Moritz S NiemiecPernilla Wittung-Stafshede
Jul 28, 2015·International Journal of Molecular Sciences·Helena Öhrvik, Pernilla Wittung-Stafshede
Jan 23, 2016·Biochemical and Biophysical Research Communications·Tanumoy MondolPernilla Wittung-Stafshede
Jan 9, 2016·Biophysical Journal·Dana KahraPernilla Wittung-Stafshede
May 28, 2015·Scientific Reports·Moritz S NiemiecPernilla Wittung-Stafshede
Feb 13, 2015·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Svenja PetzoldtPernilla Wittung-Stafshede
Jun 28, 2017·Metallomics : Integrated Biometal Science·Kumaravel Ponnandai ShanmugavelPernilla Wittung-Stafshede
Oct 25, 2017·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Candan AriözPernilla Wittung-Stafshede
Nov 6, 2015·Quarterly Reviews of Biophysics·Pernilla Wittung-Stafshede
Jul 20, 2019·Metallomics : Integrated Biometal Science·Kumaravel Ponnandai Shanmugavel, Pernilla Wittung-Stafshede
Oct 11, 2019·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Kumaravel Ponnandai ShanmugavelPernilla Wittung-Stafshede
Oct 17, 2016·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Ranjeet KumarPernilla Wittung-Stafshede
Jan 1, 2018·Quarterly Reviews of Biophysics·Candan Ariöz, Pernilla Wittung-Stafshede
Jan 1, 2018·Quarterly Reviews of Biophysics·Istvan HorvathPernilla Wittung-Stafshede
Jun 24, 2020·Biomolecules·Nima SasanianElin K Esbjörner
Nov 25, 2021·Dalton Transactions : an International Journal of Inorganic Chemistry·Radosław Kotuniak, Wojciech Bal

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Methods Mentioned

BETA
NMR
size exclusion chromatography

Software Mentioned

Matlab
Topspin
Biacore

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