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A sulfur-based transport pathway in Cu+-ATPases

Mattle, Daniel; Zhang, Limei; Sitsel, Oleg; Pedersen, Lotte Thue; Moncelli, Maria Rosa; Tadini-Buoninsegni, Francesco; Gourdon, Pontus LU ; Rees, Douglas C; Nissen, Poul and Meloni, Gabriele (2015) In EMBO Reports 16(6). p.40-728
Abstract

Cells regulate copper levels tightly to balance the biogenesis and integrity of copper centers in vital enzymes against toxic levels of copper. PIB -type Cu(+)-ATPases play a central role in copper homeostasis by catalyzing the selective translocation of Cu(+) across cellular membranes. Crystal structures of a copper-free Cu(+)-ATPase are available, but the mechanism of Cu(+) recognition, binding, and translocation remains elusive. Through X-ray absorption spectroscopy, ATPase activity assays, and charge transfer measurements on solid-supported membranes using wild-type and mutant forms of the Legionella pneumophila Cu(+)-ATPase (LpCopA), we identify a sulfur-lined metal transport pathway. Structural analysis indicates that Cu(+) is... (More)

Cells regulate copper levels tightly to balance the biogenesis and integrity of copper centers in vital enzymes against toxic levels of copper. PIB -type Cu(+)-ATPases play a central role in copper homeostasis by catalyzing the selective translocation of Cu(+) across cellular membranes. Crystal structures of a copper-free Cu(+)-ATPase are available, but the mechanism of Cu(+) recognition, binding, and translocation remains elusive. Through X-ray absorption spectroscopy, ATPase activity assays, and charge transfer measurements on solid-supported membranes using wild-type and mutant forms of the Legionella pneumophila Cu(+)-ATPase (LpCopA), we identify a sulfur-lined metal transport pathway. Structural analysis indicates that Cu(+) is bound at a high-affinity transmembrane-binding site in a trigonal-planar coordination with the Cys residues of the conserved CPC motif of transmembrane segment 4 (C382 and C384) and the conserved Met residue of transmembrane segment 6 (M717 of the MXXXS motif). These residues are also essential for transport. Additionally, the studies indicate essential roles of other conserved intramembranous polar residues in facilitating copper binding to the high-affinity site and subsequent release through the exit pathway.

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published
subject
keywords
Adenosine Triphosphatases, Amino Acid Motifs, Binding Sites, Biological Transport, Cation Transport Proteins, Cell Membrane, Copper, Legionella pneumophila, Mutagenesis, Site-Directed, Protein Binding, Protein Structure, Tertiary, Sulfur, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't
in
EMBO Reports
volume
16
issue
6
pages
13 pages
publisher
Nature Publishing Group
external identifiers
  • scopus:84930373995
ISSN
1469-221X
DOI
10.15252/embr.201439927
language
English
LU publication?
yes
id
2895781e-f0f8-4b8f-a3c7-1bc472416b11
date added to LUP
2017-04-29 15:28:15
date last changed
2017-10-01 05:33:43
@article{2895781e-f0f8-4b8f-a3c7-1bc472416b11,
  abstract     = {<p>Cells regulate copper levels tightly to balance the biogenesis and integrity of copper centers in vital enzymes against toxic levels of copper. PIB -type Cu(+)-ATPases play a central role in copper homeostasis by catalyzing the selective translocation of Cu(+) across cellular membranes. Crystal structures of a copper-free Cu(+)-ATPase are available, but the mechanism of Cu(+) recognition, binding, and translocation remains elusive. Through X-ray absorption spectroscopy, ATPase activity assays, and charge transfer measurements on solid-supported membranes using wild-type and mutant forms of the Legionella pneumophila Cu(+)-ATPase (LpCopA), we identify a sulfur-lined metal transport pathway. Structural analysis indicates that Cu(+) is bound at a high-affinity transmembrane-binding site in a trigonal-planar coordination with the Cys residues of the conserved CPC motif of transmembrane segment 4 (C382 and C384) and the conserved Met residue of transmembrane segment 6 (M717 of the MXXXS motif). These residues are also essential for transport. Additionally, the studies indicate essential roles of other conserved intramembranous polar residues in facilitating copper binding to the high-affinity site and subsequent release through the exit pathway.</p>},
  author       = {Mattle, Daniel and Zhang, Limei and Sitsel, Oleg and Pedersen, Lotte Thue and Moncelli, Maria Rosa and Tadini-Buoninsegni, Francesco and Gourdon, Pontus and Rees, Douglas C and Nissen, Poul and Meloni, Gabriele},
  issn         = {1469-221X},
  keyword      = {Adenosine Triphosphatases,Amino Acid Motifs,Binding Sites,Biological Transport,Cation Transport Proteins,Cell Membrane,Copper,Legionella pneumophila,Mutagenesis, Site-Directed,Protein Binding,Protein Structure, Tertiary,Sulfur,Journal Article,Research Support, N.I.H., Extramural,Research Support, Non-U.S. Gov't},
  language     = {eng},
  number       = {6},
  pages        = {40--728},
  publisher    = {Nature Publishing Group},
  series       = {EMBO Reports},
  title        = {A sulfur-based transport pathway in Cu+-ATPases},
  url          = {http://dx.doi.org/10.15252/embr.201439927},
  volume       = {16},
  year         = {2015},
}