The N-terminal domains of Bacillus subtilis CopA do not form a stable complex in the absence of their inter-domain linker

Zhou, Liang, Kay, Kristine L., Hecht, Oliver, Moore, Geoffrey R. and Le Brun, Nick E. ORCID: https://orcid.org/0000-0001-9780-4061 (2018) The N-terminal domains of Bacillus subtilis CopA do not form a stable complex in the absence of their inter-domain linker. Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics, 1866 (2). pp. 275-282.

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Abstract

Copper-transporting P-type ATPases, which play important roles in trafficking Cu(I) across membranes for the biogenesis of copper proteins or for copper detoxification, contain a variable number of soluble metal-binding domains at their N-termini. It is increasingly apparent that these play an important role in regulating copper transport in a Cu(I)-responsive manner, but how they do this is unknown. CopA, a Cu(I)-transporter from Bacillus subtilis, contains two N-terminal soluble domains that are closely packed, with inter-domain interactions at two principal regions. Here, we sought to determine the extent to which the domains interact in the absence of their inter-domain covalent linker, and how their Cu(I)-binding properties are affected. Studies of a 1:1 mixture of separate CopAa and CopAb domains showed that the domains do not form a stable complex, with only indirect evidence of a weak interaction between them. Their Cu(I)-binding behaviour was distinct from that of the two domain protein and consistent with a lack of interaction between the domains. Cu(I)-mediated protein association was observed, but this occurred only between domains of the same type. Thus, the inter-domain covalent link between CopAa and CopAb is essential for inter-domain interactions and for Cu(I)-binding behaviour.

Item Type:	Article
Uncontrolled Keywords:	copper trafficking,metallochaperones,homeostasis,metal transport
Faculty \ School:	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 Centres > Centre for Molecular and Structural Biochemistry Faculty of Science > Research Groups > Chemistry of Life Processes
Depositing User:	Pure Connector
Date Deposited:	15 Nov 2017 06:07
Last Modified:	30 Sep 2024 00:15
URI:	https://ueaeprints.uea.ac.uk/id/eprint/65453
DOI:	10.1016/j.bbapap.2017.11.008

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