2xla

Publication Abstract from PubMed

A copper-trafficking pathway was found to enable Cu(2+) occupancy of a soluble periplasm protein, CucA, even when competing Zn(2+) is abundant in the periplasm. Here, we solved the structure of CucA (a new cupin) and found that binding of Cu(2+), but not Zn(2+), quenches the fluorescence of Trp(165), which is adjacent to the metal site. Using this fluorescence probe, we established that CucA becomes partly occupied by Zn(2+) following exposure to equimolar Zn(2+) and Cu(2+). Cu(2+)-CucA is more thermodynamically stable than Zn(2+)-CucA but k((Zn-->Cu)exchange) is slow, raising questions about how the periplasm contains solely the Cu(2+) form. We discovered that a copper-trafficking pathway involving two copper transporters (CtaA and PacS) and a metallochaperone (Atx1) is obligatory for Cu(2+)-CucA to accumulate in the periplasm. There was negligible CucA protein in the periplasm of DeltactaA cells, but the abundance of cucA transcripts was unaltered. Crucially, DeltactaA cells overaccumulate low M(r) copper complexes in the periplasm, and purified apoCucA can readily acquire Cu(2+) from DeltactaA periplasm extracts, but in vivo apoCucA fails to come into contact with these periplasmic copper pools. Instead, copper traffics via a cytoplasmic pathway that is coupled to CucA translocation to the periplasm.

Structure and metal loading of a soluble periplasm cuproprotein.,Waldron KJ, Firbank SJ, Dainty SJ, Perez-Rama M, Tottey S, Robinson NJ J Biol Chem. 2010 Oct 15;285(42):32504-11. Epub 2010 Aug 10. PMID:20702411^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.