The N-terminal soluble domains of Bacillus subtilis CopA exhibit a high affinity and capacity for Cu(i) ions

Singleton, Chloe and Le Brun, Nick ORCID: https://orcid.org/0000-0001-9780-4061 (2009) The N-terminal soluble domains of Bacillus subtilis CopA exhibit a high affinity and capacity for Cu(i) ions. Dalton Transactions (4). pp. 688-696. ISSN 1477-9234

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Abstract

CopA from Bacillus subtilis is a Cu(I)-transporting P-type ATPase involved in resistance to high levels of environmental copper. At its N-terminus are two soluble domains, a and b, that, when generated in isolation from the membrane part, have previously been shown to exhibit unusual Cu(I)-binding behaviour: at >1 Cu(I) per CopAab the protein dimerises, resulting in the formation of a species with luminescence properties characteristic of a solvent-shielded Cu(I) cluster. Further insight into the Cu(I)-binding properties of CopAab are now reported. We demonstrate that the initial binding of Cu(I) occurs with very high affinity (K = ~ 4 × 1017 M-1) and that CopAab can accommodate up to 4 Cu(I) per protein and remains dimeric at higher Cu(I)-loadings. Fitting of UV-visible, near UV CD, fluorescence and luminescence spectroscopic titration data supports a model in which Cu(I) binds sequentially to CopAab, and also provides estimates of the association constants for Cu(I)-binding and dimerisation steps. Finally, low molecular weight thiols are shown not to affect the initial binding of Cu(I), but significantly influence binding at levels >1 Cu(I) per CopAab such that dimerisation is inhibited, though not abolished.

Item Type:	Article
Faculty \ School:	Faculty of Science > School of Chemical Sciences and Pharmacy (former - to 2009) Faculty of Science > School of Chemistry (former - to 2024)
UEA Research Groups:	Faculty of Science > Research Groups > Biophysical Chemistry (former - to 2017) Faculty of Science > Research Groups > Centre for Ocean and Atmospheric Sciences Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry Faculty of Science > Research Groups > Chemistry of Life Processes
Depositing User:	Rachel Smith
Date Deposited:	23 Mar 2011 10:35
Last Modified:	24 Sep 2024 09:25
URI:	https://ueaeprints.uea.ac.uk/id/eprint/26976
DOI:	10.1039/B810412C

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