2rml | pdb_00002rml
Solution structure of the N-terminal soluble domains of Bacillus subtilis CopA
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Overview
CopA, a P-type ATPase from Bacillus subtilis, plays a major role in the resistance of the cell to copper by effecting the export of the metal across the cytoplasmic membrane. The N-terminus of the protein features two soluble domains (a and b), that each contain a Cu(I)-binding motif, MTCAAC. We have generated a stable form of the wild-type two domain protein, CopAab, and determined its solution structure. This was found to be similar to that reported previously for a higher stability S46V variant, with minor differences mostly confined to the Ser46-containing beta3 strand of domain (a). Chemical shift analysis demonstrated that the two Cu(I)-binding motifs, located at different ends of the protein molecule, are both able to participate in Cu(I)-binding, and that Cu(I) is in rapid exchange between protein molecules. Surprisingly, UV-visible and fluorescence spectroscopy indicate very different modes of Cu(I)-binding below and above a level of 1 Cu(I) per protein, consistent with a major structural change occurring above 1 Cu(I)/CopAab. Analytical equilibrium centrifugation and gel filtration data show that this is a result of Cu(I)-mediated dimerization of the protein. The resulting species is highly luminescent, indicating the presence of a solvent-shielded Cu(I)-cluster.
About this Structure
2RML is a Single protein structure of sequence from Bacillus subtilis. Full crystallographic information is available from OCA.
Reference
Structure and Cu(I)-binding properties of the N-terminal soluble domains of Bacillus subtilis CopA., Singleton C, Banci L, Ciofi-Baffoni S, Tenori L, Kihlken M, Boetzel R, Le Brun N, Biochem J. 2008 Jan 23;. PMID:18215122
Page seeded by OCA on Wed Feb 27 07:59:27 2008
Proteopedia Page Contributors and Editors (what is this?)
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- Bacillus subtilis
- Single protein
- Banci, L.
- Bertini, I.
- Boetzel, R.
- Brun, N E.Le.
- Ciofi-Baffoni, S.
- Kihlken, M A.
- Singleton, C.
- Tenori, L.
- Atp-binding
- Copa
- Copper
- Copper transport
- Hydrolase
- Ion transport
- Magnesium
- Membrane
- Metal-binding
- Nucleotide-binding
- P-type atpase
- Phosphorylation
- Transmembrane
- Transport
