title: Rational design of true monomeric and bright photoactivatable fluorescent proteins.
abstract: Monomeric (m)Eos2 is an engineered photoactivatable fluorescent protein widely used for super-resolution microscopy. We show that mEos2 forms oligomers at high concentrations and forms aggregates when labeling membrane proteins, limiting its application as a fusion partner. We solved the crystal structure of tetrameric mEos2 and rationally designed improved versions, mEos3.1 and mEos3.2, that are truly monomeric, are brighter, mature faster and exhibit higher photon budget and label density.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
authors.forename: Mingshu
authors.initials: M
authors.lastname: Zhang
authors.forename: Hao
authors.initials: H
authors.lastname: Chang
authors.forename: Yongdeng
authors.initials: Y
authors.lastname: Zhang
authors.forename: Junwei
authors.initials: J
authors.lastname: Yu
authors.forename: Lijie
authors.initials: L
authors.lastname: Wu
authors.forename: Wei
authors.initials: W
authors.lastname: Ji
authors.forename: Juanjuan
authors.initials: J
authors.lastname: Chen
authors.forename: Bei
authors.initials: B
authors.lastname: Liu
authors.forename: Jingze
authors.initials: J
authors.lastname: Lu
authors.forename: Yingfang
authors.initials: Y
authors.lastname: Liu
authors.forename: Junlong
authors.initials: J
authors.lastname: Zhang
authors.forename: Pingyong
authors.initials: P
authors.lastname: Xu
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D002850:Chromatography, Gel; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D008958:Models, Molecular; D055668:Photochemical Processes; D010957:Plasmids; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein