PcoB is a defense outer membrane protein that facilitates cellular uptake of copper
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Dokumenter
- Fulltext
Forlagets udgivne version, 6,11 MB, PDF-dokument
Copper (Cu) is one of the most abundant trace metals in all organisms, involved in a plethora of cellular processes. Yet elevated concentrations of the element are harmful, and interestingly prokaryotes are more sensitive for environmental Cu stress than humans. Various transport systems are present to maintain intracellular Cu homeostasis, including the prokaryotic plasmid-encoded multiprotein pco operon, which is generally assigned as a defense mechanism against elevated Cu concentrations. Here we structurally and functionally characterize the outer membrane component of the Pco system, PcoB, recovering a 2.0 Å structure, revealing a classical β-barrel architecture. Unexpectedly, we identify a large opening on the extracellular side, linked to a considerably electronegative funnel that becomes narrower towards the periplasm, defining an ion-conducting pathway as also supported by metal binding quantification via inductively coupled plasma mass spectrometry and molecular dynamics (MD) simulations. However, the structure is partially obstructed towards the periplasmic side, and yet flux is permitted in the presence of a Cu gradient as shown by functional characterization in vitro. Complementary in vivo experiments demonstrate that isolated PcoB confers increased sensitivity towards Cu. Aggregated, our findings indicate that PcoB serves to permit Cu import. Thus, it is possible the Pco system physiologically accumulates Cu in the periplasm as a part of an unorthodox defense mechanism against metal stress. These results point to a previously unrecognized principle of maintaining Cu homeostasis and may as such also assist in the understanding and in efforts towards combatting bacterial infections of Pco-harboring pathogens.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | e4364 |
| Tidsskrift | Protein Science |
| Vol/bind | 31 |
| Udgave nummer | 7 |
| Sider (fra-til) | 1-16 |
| ISSN | 0961-8368 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
We are grateful for the staff at the Lund Protein Production Platform (LP3) for providing the 834(DE3) strain and for assistance with the early crystallization experiments. We also appreciate the help with the crystal screening and data collection at the SLS, the Paul Scherrer Institute, Villigen, beam line X06SAWe acknowledge Zhila Nikrozi and Klaus Qvortrup from the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen for the help of preparing fixed bacteria TEM sample preparation and imaging, and Prof. Michael Davies for access to stop‐flow cytometry equipment. Access to synchrotron sources was supported by The Danscatt program of The Independent Research Fund Denmark. GM is supported by the National Institute of General Medical Sciences of the National Institutes of Health and by the Robert A. Welch foundation. PG is supported by the following Foundations: Lundbeck, Knut and Alice Wallenberg, Carlsberg, Novo‐Nordisk, Brødrene Hartmann, Agnes og Poul Friis, Augustinus, Crafoord as well as The Per‐Eric and Ulla Schyberg. Funding is also obtained from The Independent Research Fund Denmark, the Swedish Research Council and through a Michaelsen scholarship. DRM was funded by Carl Tryggers foundation, MA was funded by a Swedish Research Council Starting Grant. The computations were performed on resources provided by the Swedish National Infrastructure for Computing (SNIC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. E. coli
Funding Information:
We are grateful for the staff at the Lund Protein Production Platform (LP3) for providing the E. coli 834(DE3) strain and for assistance with the early crystallization experiments. We also appreciate the help with the crystal screening and data collection at the SLS, the Paul Scherrer Institute, Villigen, beam line X06SAWe acknowledge Zhila Nikrozi and Klaus Qvortrup from the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen for the help of preparing fixed bacteria TEM sample preparation and imaging, and Prof. Michael Davies for access to stop-flow cytometry equipment. Access to synchrotron sources was supported by The Danscatt program of The Independent Research Fund Denmark. GM is supported by the National Institute of General Medical Sciences of the National Institutes of Health and by the Robert A. Welch foundation. PG is supported by the following Foundations: Lundbeck, Knut and Alice Wallenberg, Carlsberg, Novo-Nordisk, Brødrene Hartmann, Agnes og Poul Friis, Augustinus, Crafoord as well as The Per-Eric and Ulla Schyberg. Funding is also obtained from The Independent Research Fund Denmark, the Swedish Research Council and through a Michaelsen scholarship. DRM was funded by Carl Tryggers foundation, MA was funded by a Swedish Research Council Starting Grant. The computations were performed on resources provided by the Swedish National Infrastructure for Computing (SNIC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.
Antal downloads er baseret på statistik fra Google Scholar og www.ku.dk
ID: 313883710