title: Crystal structures of Dronpa complexed with quenchable metal ions provide insight into metal biosensor development.
abstract: Many fluorescent proteins (FPs) show fluorescence quenching by specific metal ions, which can be applied towards metal biosensor development. In this study, we investigated the significant fluorescence quenching of Dronpa by Co(2+) and Cu(2+) ions. Crystal structures of Co(2+) -, Ni(2+) - and Cu(2+) -bound Dronpa revealed previously unseen, unique, metal-binding sites for fluorescence quenching. These metal ions commonly interact with surface-exposed histidine residues (His194-His210 and His210-His212), and interact indirectly with chromophores. Structural analysis of the Co(2+) - and Cu(2+) - binding sites of Dronpa provides insight into FP-based metal biosensor engineering.
authors.affiliation: Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Seoul, Korea.
authors.forename: In Jung
authors.initials: IJ
authors.lastname: Kim
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Sangsoo
authors.initials: S
authors.lastname: Kim
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Jeahyun
authors.initials: J
authors.lastname: Park
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Intae
authors.initials: I
authors.lastname: Eom
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Sunam
authors.initials: S
authors.lastname: Kim
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Sung Chul
authors.initials: SC
authors.lastname: Ha
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Yeon Gil
authors.initials: YG
authors.lastname: Kim
authors.affiliation: Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Seoul, Korea.
authors.forename: Kwang Yeon
authors.initials: KY
authors.lastname: Hwang
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Korea.
authors.forename: Ki Hyun
authors.initials: KH
authors.lastname: Nam
mesh_terms: D001665:Binding Sites; D015374:Biosensing Techniques; D003035:Cobalt / Q000737:chemistry; D003300:Copper / Q000737:chemistry; D018360:Crystallography, X-Ray; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D006801:Humans; D007477:Ions / Q000737:chemistry; D008670:Metals / Q000737:chemistry / Q000302:isolation & purification*; D009532:Nickel / Q000737:chemistry; D011487:Protein Conformation
chemical_list: D007477:Ions; D008670:Metals; D049452:Green Fluorescent Proteins; D003035:Cobalt; D003300:Copper; D009532:Nickel
keywords: Dronpa; biosensor; fluorescent protein; metal-binding site; quenching