title: Structural basis for the influence of a single mutation K145N on the oligomerization and photoswitching rate of Dronpa.
abstract: The crystal structure of the on-state of PDM1-4, a single-mutation variant of the photochromic fluorescent protein Dronpa, is reported at 1.95 Å resolution. PDM1-4 is a Dronpa variant that possesses a slower off-switching rate than Dronpa and thus can effectively increase the image resolution in subdiffraction optical microscopy, although the precise molecular basis for this change has not been elucidated. This work shows that the Lys145Asn mutation in PDM1-4 stabilizes the interface available for dimerization, facilitating oligomerization of the protein. No significant changes were observed in the chromophore environment of PDM1-4 compared with Dronpa, and the ensemble absorption and emission properties of PDM1-4 were highly similar to those of Dronpa. It is proposed that the slower off-switching rate in PDM1-4 is caused by a decrease in the potential flexibility of certain β-strands caused by oligomerization along the AC interface.
authors.affiliation: Biomolecular Architecture, Department of Chemistry, KU Leuven, Celestijnenlaan 200F, Leuven (Heverlee), Belgium.
authors.forename: Ngan
authors.initials: N
authors.lastname: Nguyen Bich
authors.forename: Benjamien
authors.initials: B
authors.lastname: Moeyaert
authors.forename: Kristof
authors.initials: K
authors.lastname: Van Hecke
authors.forename: Peter
authors.initials: P
authors.lastname: Dedecker
authors.forename: Hideaki
authors.initials: H
authors.lastname: Mizuno
authors.forename: Johan
authors.initials: J
authors.lastname: Hofkens
authors.forename: Luc
authors.initials: L
authors.lastname: Van Meervelt
mesh_terms: D001704:Biopolymers / Q000737:chemistry*; D002846:Chromatography, Affinity; D018360:Crystallography, X-Ray; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D007700:Kinetics; D008027:Light; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation
chemical_list: D001704:Biopolymers; D049452:Green Fluorescent Proteins