Acquisition of ionic copper by a bacterial outer membrane protein

Bhamidimarri, S.; Young, T.; Shanmugam, M.; Soderholm, S.; Basle, A.; Belzunces, B.; Skylaris, C.; Bumann, D.; Khalid, S.; van den Berg, B.

doi:10.1101/2020.06.04.134395

bioRxiv · 10.1101/2020.06.04.134395

Acquisition of ionic copper by a bacterial outer membrane protein

Bhamidimarri, S.·Young, T.·Shanmugam, M.·Soderholm, S.·Basle, A.·Belzunces, B.·Skylaris, C.·Bumann, D.·Khalid, S.·van den Berg, B.

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Abstract

Copper, while toxic in excess, is an essential micronutrient in all kingdoms of life due to its essential role in the structure and function of many proteins. Proteins mediating ionic copper import have been characterised in detail for eukaryotes, but much less so for prokaryotes. In particular, it is still unclear whether and how Gram-negative bacteria acquire ionic copper. Here we show that Pseudomonas aeruginosa OprC is an outer membrane, TonB-dependent transporter that is conserved in many Proteobacteria and which mediates acquisition of both reduced and oxidised ionic copper via an unprecedented CxxxM-HxM metal binding site. Crystal structures of wild type and mutant OprC variants with silver and copper suggest that acquisition of Cu(I) occurs via a surface-exposed "methionine track" leading towards the principal metal binding site. Together with whole-cell copper quantitation and quantitative proteomics in a murine lung infection model, our data identify OprC as an abundant component of bacterial copper biology that may enable copper acquisition under a wide range of conditions. SignificanceCopper is an essential metal in biology due to its role in the structure and function of many proteins. Despite this, it is not very clear how bacteria acquire copper, especially for Gram-negative organisms. In this study we show that the outer membrane protein OprC has an unusual metal binding site that allows OprC to bind both reduced and oxidised ionic copper near-irreversibly. Given the versatility of OprC, its presence in many Proteobacteria and its abundance during lung infection in mice, our study shows that OprC is an important component of prokaryote copper biology that warrants further study to uncover its regulation and to assess its role in bacterial virulence.

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Bhamidimarri, S., Young, T., Shanmugam, M., Soderholm, S., Basle, A., Belzunces, B., Skylaris, C., Bumann, D., Khalid, S., van den Berg, B.. 2020-06-05. Acquisition of ionic copper by a bacterial outer membrane protein. https://doi.org/10.1101/2020.06.04.134395

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