title: Competitive cobalt for zinc substitution in mammalian methionine sulfoxide reductase B1 overexpressed in E. coli: structural and functional insight.
abstract: Expression of the mammalian enzyme methionine sulfoxide reductase B1 (MsrB1) in Escherichia coli growing in cobalt-containing media resulted in the reproducible appearance of the stable cobalt-containing protein MsrB1-Co. NMR studies and biocomputing using the programs AnisoFit and Amber allowed us to generate a structure of MsrB1-Co sharing the overall fold with the native zinc-containing protein MsrB1-Zn. Our data suggest that the N-terminus containing resolving cysteine tends to be closer to the protein's catalytic center than was previously reported. It is argued that this proximity supports the proposed catalytic mechanism and ensures high catalytic efficiency of MsrB1. Functional studies showed that both MsrB1-Zn and MsrB1-Co exhibit similar levels of activity, in agreement with the structural studies performed. The proposed metal ion substitution approach may have a methodological significance in determining whether methionine sulfoxide reductase B proteins contain a metal ion.
authors.affiliation: Department of Biotechnology, Norwegian University of Science and Technology, 7491, Trondheim, Norway.
authors.forename: Elena
authors.initials: E
authors.lastname: Shumilina
authors.forename: Olena
authors.initials: O
authors.lastname: Dobrovolska
authors.forename: Rebecca
authors.initials: R
authors.lastname: Del Conte
authors.forename: Henrik Waldal
authors.initials: HW
authors.lastname: Holen
authors.forename: Alexander
authors.initials: A
authors.lastname: Dikiy
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D003035:Cobalt / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D051150:Methionine Sulfoxide Reductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D011487:Protein Conformation; D015854:Up-Regulation; D015032:Zinc / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003035:Cobalt; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase; D015032:Zinc