title: Identification of major zinc-binding proteins from a marine cyanobacterium: insight into metal uptake in oligotrophic environments.
abstract: Marine cyanobacteria make a significant contribution to primary production whilst occupying some of the most nutrient poor regions of the world's oceans. The low bioavailability of trace metals can limit the growth of phytoplankton in ocean waters, but only scarce data are available on the requirements of marine microbes for zinc. Recent genome mining studies suggest that marine cyanobacteria have both uptake systems for zinc and proteins that utilize zinc as a cofactor. In this study, the oligotrophic strain Synechococcus sp. WH8102 was grown at different zinc concentrations. Using metalloproteomics approaches, we demonstrate that even though this organism's growth was not affected by extremely low zinc levels, cells accumulated significant quantities of zinc, which was shown to be protein-associated by 2D liquid chromatography and ICP-MS. This indicates that the mechanisms for zinc uptake in Synechococcus sp. WH8102 are extremely efficient. Significantly, expression of SYNW2224, a putative porin, was up-regulated during growth in zinc-depleted conditions. Furthermore, along with 30 other proteins, SYNW2224 was captured by immobilised zinc affinity chromatography, indicating the presence of surface-exposed site(s) with metal-binding capacity. It is proposed that this porin plays a role in high-affinity zinc uptake in this and other cyanobacteria.
authors.affiliation: Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK. c.blindauer@warwick.ac.uk.
authors.forename: James Paul
authors.initials: JP
authors.lastname: Barnett
authors.forename: David John
authors.initials: DJ
authors.lastname: Scanlan
authors.forename: Claudia Andrea
authors.initials: CA
authors.lastname: Blindauer
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D001665:Binding Sites; D002352:Carrier Proteins / Q000302:isolation & purification*; D027682:Cation Transport Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D008958:Models, Molecular; D018272:Porins / Q000302:isolation & purification*; D046940:Synechococcus; D015032:Zinc / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D027682:Cation Transport Proteins; D018272:Porins; C017800:zinc-binding protein; D015032:Zinc