Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution

Amy C. Rosenzweig, David L. Huffman, Melody Hou, Amy K. Wernimont, Robert A. Pufahl, Thomas V. O'Halloran

Research output: Contribution to journalArticle

207 Citations (Scopus)

Abstract

Background: Metallochaperone proteins function in the trafficking and delivery of essential, yet potentially toxic, metal ions to distinct locations and particular proteins in eukaryotic cells. The Atx1 protein shuttles copper to the transport ATPase Ccc2 in yeast cells. Molecular mechanisms for copper delivery by Atx1 and similar human chaperones have been proposed, but detailed structural characterization is necessary to elucidate how Atx1 binds metal ions and how it might interact with Ccc2 to facilitate metal ion transfer. Results: The 1.02 Å resolution X-ray structure of the Hg(II) form of Atx1 (HgAtx1) reveals the overall secondary structure, the location of the metal-binding site, the detailed coordination geometry for Hg(II), and specific amino acid residues that may be important in interactions with Ccc2. Metal ion transfer experiments establish that HgAtx1 is a functional model for the Cu(l) form of Atx1 (CuAtx1). The metal-binding loop is flexible, changing conformation to form a disulfide bond in the oxidized apo form, the structure of which has been solved to 1.20 Å resolution. Conclusions: The Atx1 structure represents the first structure of a metallochaperone protein, and is one of the largest unknown structures solved by direct methods. The structural features of the metal-binding site support the proposed Atx1 mechanism in which facile metal ion transfer occurs between metal-binding sites of the diffusible copper-donor and membrane- tethered copper-acceptor proteins. The Atx1 structural motif represents a prototypical metal ion trafficking unit that is likely to be employed in a variety of organisms for different metal ions.

Original languageEnglish (US)
Pages (from-to)605-617
Number of pages13
JournalStructure
Volume7
Issue number6
DOIs
StatePublished - Jun 1999
Externally publishedYes

Fingerprint

Metallochaperones
Metals
Ions
Proteins
Copper
Binding Sites
Poisons
Eukaryotic Cells
Disulfides

Keywords

  • Copper transport
  • Direct methods
  • Menkes syndrome
  • Mercury coordination
  • Shake and Bake

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

Cite this

Rosenzweig, A. C., Huffman, D. L., Hou, M., Wernimont, A. K., Pufahl, R. A., & O'Halloran, T. V. (1999). Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution. Structure, 7(6), 605-617. https://doi.org/10.1016/S0969-2126(99)80082-3

Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution. / Rosenzweig, Amy C.; Huffman, David L.; Hou, Melody; Wernimont, Amy K.; Pufahl, Robert A.; O'Halloran, Thomas V.

In: Structure, Vol. 7, No. 6, 06.1999, p. 605-617.

Research output: Contribution to journalArticle

Rosenzweig, AC, Huffman, DL, Hou, M, Wernimont, AK, Pufahl, RA & O'Halloran, TV 1999, 'Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution', Structure, vol. 7, no. 6, pp. 605-617. https://doi.org/10.1016/S0969-2126(99)80082-3
Rosenzweig AC, Huffman DL, Hou M, Wernimont AK, Pufahl RA, O'Halloran TV. Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution. Structure. 1999 Jun;7(6):605-617. https://doi.org/10.1016/S0969-2126(99)80082-3
Rosenzweig, Amy C. ; Huffman, David L. ; Hou, Melody ; Wernimont, Amy K. ; Pufahl, Robert A. ; O'Halloran, Thomas V. / Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution. In: Structure. 1999 ; Vol. 7, No. 6. pp. 605-617.
@article{d7deac73f21e4d08b71bf1d8b46afbfd,
title = "Crystal structure of the Atx1 metallochaperone protein at 1.02 {\AA} resolution",
abstract = "Background: Metallochaperone proteins function in the trafficking and delivery of essential, yet potentially toxic, metal ions to distinct locations and particular proteins in eukaryotic cells. The Atx1 protein shuttles copper to the transport ATPase Ccc2 in yeast cells. Molecular mechanisms for copper delivery by Atx1 and similar human chaperones have been proposed, but detailed structural characterization is necessary to elucidate how Atx1 binds metal ions and how it might interact with Ccc2 to facilitate metal ion transfer. Results: The 1.02 {\AA} resolution X-ray structure of the Hg(II) form of Atx1 (HgAtx1) reveals the overall secondary structure, the location of the metal-binding site, the detailed coordination geometry for Hg(II), and specific amino acid residues that may be important in interactions with Ccc2. Metal ion transfer experiments establish that HgAtx1 is a functional model for the Cu(l) form of Atx1 (CuAtx1). The metal-binding loop is flexible, changing conformation to form a disulfide bond in the oxidized apo form, the structure of which has been solved to 1.20 {\AA} resolution. Conclusions: The Atx1 structure represents the first structure of a metallochaperone protein, and is one of the largest unknown structures solved by direct methods. The structural features of the metal-binding site support the proposed Atx1 mechanism in which facile metal ion transfer occurs between metal-binding sites of the diffusible copper-donor and membrane- tethered copper-acceptor proteins. The Atx1 structural motif represents a prototypical metal ion trafficking unit that is likely to be employed in a variety of organisms for different metal ions.",
keywords = "Copper transport, Direct methods, Menkes syndrome, Mercury coordination, Shake and Bake",
author = "Rosenzweig, {Amy C.} and Huffman, {David L.} and Melody Hou and Wernimont, {Amy K.} and Pufahl, {Robert A.} and O'Halloran, {Thomas V.}",
year = "1999",
month = "6",
doi = "10.1016/S0969-2126(99)80082-3",
language = "English (US)",
volume = "7",
pages = "605--617",
journal = "Structure with Folding & design",
issn = "0969-2126",
publisher = "Cell Press",
number = "6",

}

TY - JOUR

T1 - Crystal structure of the Atx1 metallochaperone protein at 1.02 Å resolution

AU - Rosenzweig, Amy C.

AU - Huffman, David L.

AU - Hou, Melody

AU - Wernimont, Amy K.

AU - Pufahl, Robert A.

AU - O'Halloran, Thomas V.

PY - 1999/6

Y1 - 1999/6

N2 - Background: Metallochaperone proteins function in the trafficking and delivery of essential, yet potentially toxic, metal ions to distinct locations and particular proteins in eukaryotic cells. The Atx1 protein shuttles copper to the transport ATPase Ccc2 in yeast cells. Molecular mechanisms for copper delivery by Atx1 and similar human chaperones have been proposed, but detailed structural characterization is necessary to elucidate how Atx1 binds metal ions and how it might interact with Ccc2 to facilitate metal ion transfer. Results: The 1.02 Å resolution X-ray structure of the Hg(II) form of Atx1 (HgAtx1) reveals the overall secondary structure, the location of the metal-binding site, the detailed coordination geometry for Hg(II), and specific amino acid residues that may be important in interactions with Ccc2. Metal ion transfer experiments establish that HgAtx1 is a functional model for the Cu(l) form of Atx1 (CuAtx1). The metal-binding loop is flexible, changing conformation to form a disulfide bond in the oxidized apo form, the structure of which has been solved to 1.20 Å resolution. Conclusions: The Atx1 structure represents the first structure of a metallochaperone protein, and is one of the largest unknown structures solved by direct methods. The structural features of the metal-binding site support the proposed Atx1 mechanism in which facile metal ion transfer occurs between metal-binding sites of the diffusible copper-donor and membrane- tethered copper-acceptor proteins. The Atx1 structural motif represents a prototypical metal ion trafficking unit that is likely to be employed in a variety of organisms for different metal ions.

AB - Background: Metallochaperone proteins function in the trafficking and delivery of essential, yet potentially toxic, metal ions to distinct locations and particular proteins in eukaryotic cells. The Atx1 protein shuttles copper to the transport ATPase Ccc2 in yeast cells. Molecular mechanisms for copper delivery by Atx1 and similar human chaperones have been proposed, but detailed structural characterization is necessary to elucidate how Atx1 binds metal ions and how it might interact with Ccc2 to facilitate metal ion transfer. Results: The 1.02 Å resolution X-ray structure of the Hg(II) form of Atx1 (HgAtx1) reveals the overall secondary structure, the location of the metal-binding site, the detailed coordination geometry for Hg(II), and specific amino acid residues that may be important in interactions with Ccc2. Metal ion transfer experiments establish that HgAtx1 is a functional model for the Cu(l) form of Atx1 (CuAtx1). The metal-binding loop is flexible, changing conformation to form a disulfide bond in the oxidized apo form, the structure of which has been solved to 1.20 Å resolution. Conclusions: The Atx1 structure represents the first structure of a metallochaperone protein, and is one of the largest unknown structures solved by direct methods. The structural features of the metal-binding site support the proposed Atx1 mechanism in which facile metal ion transfer occurs between metal-binding sites of the diffusible copper-donor and membrane- tethered copper-acceptor proteins. The Atx1 structural motif represents a prototypical metal ion trafficking unit that is likely to be employed in a variety of organisms for different metal ions.

KW - Copper transport

KW - Direct methods

KW - Menkes syndrome

KW - Mercury coordination

KW - Shake and Bake

UR - http://www.scopus.com/inward/record.url?scp=0033153380&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033153380&partnerID=8YFLogxK

U2 - 10.1016/S0969-2126(99)80082-3

DO - 10.1016/S0969-2126(99)80082-3

M3 - Article

C2 - 10404590

AN - SCOPUS:0033153380

VL - 7

SP - 605

EP - 617

JO - Structure with Folding & design

JF - Structure with Folding & design

SN - 0969-2126

IS - 6

ER -