Abstract
Copper cations play fundamental roles in biological systems, such as protein folding and stabilization, or enzymatic reactions. Although copper is essential to the cell, it can become cytotoxic if present in too high concentration. Organisms have therefore developed specific regulation mechanisms towards copper. This is the case of the Pco system present in the bacterium Caulobacter crescentus, which is composed of two proteins: a soluble periplasmic protein PcoA and an outer membrane protein PcoB. PcoA oxidizes Cu + to Cu 2+, whereas PcoB is thought to be an efflux pump for Cu 2+. While the PcoA protein has already been studied, very little is known about the structure and function of PcoB. In the present work, PcoB has been overexpressed in high yield in E. coli strains and successfully refolded by the SDS-cosolvent method. Binding to divalent cations has also been studied using several spectroscopic techniques. In addition, a three-dimensional structure model of PcoB, experimentally supported by circular dichroism, has been constructed, showing a β-barrel conformation with a N-terminal disordered chain. This peculiar intrinsic disorder property has also been confirmed by various bioinformatic tools.
Original language | English |
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Article number | 184038 |
Journal | Biochimica et Biophysica Acta - Biomembranes |
Volume | 1864 |
Issue number | 12 |
DOIs | |
Publication status | Published - 1 Dec 2022 |
Keywords
- Caulobacter crescentus
- Copper-binding protein
- Intrinsically disordered membrane protein
- PcoB
- Refolding
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Student theses
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Characterization of the molecular role of PcoB in copper efflux in Caulobacter crescentus
Author: Khochtali, A., 12 Feb 2024Supervisor: Matroule, J. (Supervisor), De Bolle, X. (President), Michaux, C. (Jury), Jacob-Dubuisson, F. (External person) (Jury) & Ilbert, M. (External person) (Jury)
Student thesis: Doc types › Doctor of Sciences