title: Histidine ligands in bacterial metallothionein enhance cluster stability.
abstract: The cyanobacterial metallothionein (MT) SmtA is the prototype for bacterial MTs and protects against elevated levels of zinc. In contrast to mammalian MTs, bacterial MTs coordinate to metal ions not only via cysteine sulfurs, but unusually for MTs, also via histidine nitrogens. To investigate whether histidine coordination in these metal-sulfur clusters provides advantages over S-coordination only, we mutated the two metal-binding histidine residues in the cyanobacterial MT SmtA from Synechococcus PCC7942 to cysteines. We show that the mutant proteins are still capable of binding up to four zinc ions as is the wild-type protein. However, the mutations perturb protein folding and metal-binding dynamics. Interestingly, several homologues of SmtA also show variations in these two residues. We conclude that histidine residues in Synechococcus PCC7942 SmtA have a stabilising effect due to electrostatic interactions that impact on protein folding and metal cluster charge, and are involved in fine-tuning the reactivity of the bound metal ions.
authors.affiliation: School of Chemistry, University of Edinburgh, West Mains Road, Edinburgh, EH9 3JJ, UK. c.blindauer@warwick.ac.uk
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Blindauer
authors.forename: M Tahir
authors.initials: MT
authors.lastname: Razi
authors.forename: Dominic J
authors.initials: DJ
authors.lastname: Campopiano
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Sadler
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002104:Cadmium / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry / Q000235:genetics; D006639:Histidine / Q000737:chemistry* / Q000235:genetics; D008024:Ligands; D008668:Metallothionein / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011487:Protein Conformation; D017510:Protein Folding; D017386:Sequence Homology, Amino Acid; D055672:Static Electricity; D015032:Zinc / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; C080048:smtA protein, Synechococcus; D002104:Cadmium; D006639:Histidine; D008668:Metallothionein; D015032:Zinc; D003545:Cysteine