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Title: The cbb 3-type cytochrome oxidase assembly factor CcoG is a widely distributed cupric reductase

Abstract

Copper (Cu)-containing proteins execute essential functions in prokaryotic and eukaryotic cells, but their biogenesis is challenged by high Cu toxicity and the preferential presence of Cu(II) under aerobic conditions, while Cu(I) is the preferred substrate for Cu chaperones and Cu-transport proteins. These proteins form a coordinated network that prevents Cu accumulation, which would lead to toxic effects such as Fenton-like reactions and mismetalation of other metalloproteins. Simultaneously, Cu-transport proteins and Cu chaperones sustain Cu(I) supply for cuproprotein biogenesis and are therefore essential for the biogenesis of Cu-containing proteins. In eukaryotes, Cu(I) is supplied for import and trafficking by cell-surface exposed metalloreductases, but specific cupric reductases have not been identified in bacteria. It was generally assumed that the reducing environment of the bacterial cytoplasm would suffice to provide sufficient Cu(I) for detoxification and cuproprotein synthesis. Here, we identify the proposed cbb 3 -type cytochrome c oxidase ( cbb 3 -Cox) assembly factor CcoG as a cupric reductase that binds Cu via conserved cysteine motifs and contains 2 low-potential [4Fe-4S] clusters required for Cu(II) reduction. Deletion of ccoG or mutation of the cysteine residues results in defective cbb 3 -Cox assembly and Cu sensitivity. Furthermore, anaerobically purified CcoG catalyzes Cu(II) but notmore » Fe(III) reduction in vitro using an artificial electron donor. Thus, CcoG is a bacterial cupric reductase and a founding member of a widespread class of enzymes that generate Cu(I) in the bacterial cytosol by using [4Fe-4S] clusters.« less

Authors:
 [1];  [2];  [1];  [2];  [1];  [3]; ORCiD logo [4];  [1];  [2]; ORCiD logo [1]
  1. Albert Ludwigs Univ. of Freiburg, Freiburg (Germany)
  2. Univ. of Pennsylvania, Philadelphia, PA (United States)
  3. Univ. of California, Berkeley, CA (United States)
  4. Brookhaven National Lab. (BNL), Upton, NY (United States)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Chemical Sciences, Geosciences & Biosciences Division
OSTI Identifier:
1579892
Alternate Identifier(s):
OSTI ID: 1567869
Report Number(s):
BNL-212441-2019-JAAM
Journal ID: ISSN 0027-8424
Grant/Contract Number:  
SC0012704; FG02-91ER20052; GM 38237; GM42143
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 42; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; cytochrome oxidase; copper; membrane protein; biogenesis; Rhodobacter capsulatus; homeostasis

Citation Formats

Marckmann, Dorian, Trasnea, Petru -Iulian, Schimpf, Johannes, Winterstein, Christine, Andrei, Andreea, Schmollinger, Stefan, Blaby-Haas, Crysten E., Friedrich, Thorsten, Daldal, Fevzi, and Koch, Hans -Georg. The cbb3-type cytochrome oxidase assembly factor CcoG is a widely distributed cupric reductase. United States: N. p., 2019. Web. doi:10.1073/pnas.1913803116.
Marckmann, Dorian, Trasnea, Petru -Iulian, Schimpf, Johannes, Winterstein, Christine, Andrei, Andreea, Schmollinger, Stefan, Blaby-Haas, Crysten E., Friedrich, Thorsten, Daldal, Fevzi, & Koch, Hans -Georg. The cbb3-type cytochrome oxidase assembly factor CcoG is a widely distributed cupric reductase. United States. doi:10.1073/pnas.1913803116.
Marckmann, Dorian, Trasnea, Petru -Iulian, Schimpf, Johannes, Winterstein, Christine, Andrei, Andreea, Schmollinger, Stefan, Blaby-Haas, Crysten E., Friedrich, Thorsten, Daldal, Fevzi, and Koch, Hans -Georg. Mon . "The cbb3-type cytochrome oxidase assembly factor CcoG is a widely distributed cupric reductase". United States. doi:10.1073/pnas.1913803116.
@article{osti_1579892,
title = {The cbb3-type cytochrome oxidase assembly factor CcoG is a widely distributed cupric reductase},
author = {Marckmann, Dorian and Trasnea, Petru -Iulian and Schimpf, Johannes and Winterstein, Christine and Andrei, Andreea and Schmollinger, Stefan and Blaby-Haas, Crysten E. and Friedrich, Thorsten and Daldal, Fevzi and Koch, Hans -Georg},
abstractNote = {Copper (Cu)-containing proteins execute essential functions in prokaryotic and eukaryotic cells, but their biogenesis is challenged by high Cu toxicity and the preferential presence of Cu(II) under aerobic conditions, while Cu(I) is the preferred substrate for Cu chaperones and Cu-transport proteins. These proteins form a coordinated network that prevents Cu accumulation, which would lead to toxic effects such as Fenton-like reactions and mismetalation of other metalloproteins. Simultaneously, Cu-transport proteins and Cu chaperones sustain Cu(I) supply for cuproprotein biogenesis and are therefore essential for the biogenesis of Cu-containing proteins. In eukaryotes, Cu(I) is supplied for import and trafficking by cell-surface exposed metalloreductases, but specific cupric reductases have not been identified in bacteria. It was generally assumed that the reducing environment of the bacterial cytoplasm would suffice to provide sufficient Cu(I) for detoxification and cuproprotein synthesis. Here, we identify the proposed cbb 3 -type cytochrome c oxidase ( cbb 3 -Cox) assembly factor CcoG as a cupric reductase that binds Cu via conserved cysteine motifs and contains 2 low-potential [4Fe-4S] clusters required for Cu(II) reduction. Deletion of ccoG or mutation of the cysteine residues results in defective cbb 3 -Cox assembly and Cu sensitivity. Furthermore, anaerobically purified CcoG catalyzes Cu(II) but not Fe(III) reduction in vitro using an artificial electron donor. Thus, CcoG is a bacterial cupric reductase and a founding member of a widespread class of enzymes that generate Cu(I) in the bacterial cytosol by using [4Fe-4S] clusters.},
doi = {10.1073/pnas.1913803116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 42,
volume = 116,
place = {United States},
year = {2019},
month = {9}
}

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