Conformations of the apo-, substrate-bound and phosphate-bound ATP-binding domain of the Cu(II) ATPase CopB illustrate coupling of domain movement to the catalytic cycle

Samuel Jayakanthan, Sue A Roberts, Andrzej Weichsel, José M. Argüello, Megan McEvoy

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Heavy metal P1B-type ATPases play a critical role in cell survival by maintaining appropriate intracellular metal concentrations. Archaeoglobus fulgidus CopB is a member of this family that transports Cu(II) from the cytoplasm to the exterior of the cell using ATP as energy source. CopB has a 264 amino acid ATPBD (ATP-binding domain) that is essential for ATP binding and hydrolysis as well as ultimately transducing the energy to the transmembrane metal-binding site for metal occlusion and export. The relevant conformations of this domain during the different steps of the catalytic cycle are still under discussion. Through crystal structures of the apo- and phosphate-bound ATPBDs, with limited proteolysis and fluorescence studies of the apo- and substrate-bound states, we show that the isolated ATPBD of CopB cycles from an open conformation in the apo-state to a closed conformation in the substrate-bound state, then returns to an open conformation suitable for product release. The present work is the first structural report of an ATPBD with its physiologically relevant product (phosphate) bound. The solution studies we have performed help resolve questions on the potential influence of crystal packing on domain conformation. These results explain how phosphate is co-ordinated in ATPase transporters and give an insight into the physiologically relevant conformation of the ATPBD at different steps of the catalytic cycle.

Original languageEnglish (US)
Pages (from-to)443-453
Number of pages11
JournalBioscience Reports
Volume32
Issue number5
DOIs
StatePublished - Oct 2012

Fingerprint

Adenosine Triphosphatases
Conformations
Adenosine Triphosphate
Phosphates
Substrates
Metals
Archaeoglobus fulgidus
Proteolysis
Heavy Metals
Hydrolysis
Cell Survival
Cytoplasm
Crystal structure
Fluorescence
Binding Sites
Cells
Amino Acids
Crystals

Keywords

  • ATP-binding domain (ATPBD)
  • ATPase transporter
  • CopB
  • Copper
  • Crystal structure
  • Metal transport

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Conformations of the apo-, substrate-bound and phosphate-bound ATP-binding domain of the Cu(II) ATPase CopB illustrate coupling of domain movement to the catalytic cycle. / Jayakanthan, Samuel; Roberts, Sue A; Weichsel, Andrzej; Argüello, José M.; McEvoy, Megan.

In: Bioscience Reports, Vol. 32, No. 5, 10.2012, p. 443-453.

Research output: Contribution to journalArticle

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