title: Cyanobacterial metallochaperone inhibits deleterious side reactions of copper.
abstract: Copper metallochaperones supply copper to cupro-proteins through copper-mediated protein-protein-interactions and it has been hypothesized that metallochaperones thereby inhibit copper from causing damage en route. Evidence is presented in support of this latter role for cyanobacterial metallochaperone, Atx1. In cyanobacteria Atx1 contributes towards the supply of copper to plastocyanin inside thylakoids but it is shown here that in copper-replete medium, copper can reach plastocyanin without Atx1. Unlike metallochaperone-independent copper-supply to superoxide dismutase in eukaryotes, glutathione is not essential for Atx1-independent supply to plastocyanin: Double mutants missing atx1 and gshB (encoding glutathione synthetase) accumulate the same number of atoms of copper per cell in the plastocyanin pool as wild type. Critically, Δatx1ΔgshB are hypersensitive to elevated copper relative to wild type cells and also relative to ΔgshB single mutants with evidence that hypersensitivity arises due to the mislocation of copper to sites for other metals including iron and zinc. The zinc site on the amino-terminal domain (ZiaA(N)) of the P(1)-type zinc-transporting ATPase is especially similar to the copper site of the Atx1 target PacS(N), and ZiaA(N) will bind Cu(I) more tightly than zinc. An NMR model of a substituted-ZiaA(N)-Cu(I)-Atx1 heterodimer has been generated making it possible to visualize a juxtaposition of residues surrounding the ZiaA(N) zinc site, including Asp(18), which normally repulse Atx1. Equivalent repulsion between bacterial copper metallochaperones and the amino-terminal regions of P(1)-type ATPases for metals other than Cu(I) is conserved, again consistent with a role for copper metallochaperones to withhold copper from binding sites for other metals.
authors.affiliation: Department of Chemistry, Biophysical Sciences Institute, School of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, United Kingdom.
authors.forename: Steve
authors.initials: S
authors.lastname: Tottey
authors.forename: Carl J
authors.initials: CJ
authors.lastname: Patterson
authors.forename: Lucia
authors.initials: L
authors.lastname: Banci
authors.forename: Ivano
authors.initials: I
authors.lastname: Bertini
authors.forename: Isabella C
authors.initials: IC
authors.lastname: Felli
authors.forename: Anna
authors.initials: A
authors.lastname: Pavelkova
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Dainty
authors.forename: Rafael
authors.initials: R
authors.lastname: Pernil
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Waldron
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Foster
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Robinson
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D003300:Copper / Q000494:pharmacology / Q000633:toxicity*; D003470:Culture Media / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006706:Homeostasis / Q000187:drug effects; D056511:Metallochaperones / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D010970:Plastocyanin / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D046939:Synechocystis / Q000187:drug effects* / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D056511:Metallochaperones; D003300:Copper; D010970:Plastocyanin; D000251:Adenosine Triphosphatases; D005978:Glutathione; D015032:Zinc