title: mBeRFP, an improved large stokes shift red fluorescent protein.
abstract: Herein, we describe the generation of a monomeric large Stokes shift (LSS) red fluorescent protein, mBeRFP, with excitation and emission peaks at 446 and 615 nm, respectively. Compared with two previously reported LSS-RFPs (mKeima and LSS-mKate2), mBeRFP is approximately three times brighter. In addition, mBeRFP is characterized by improved photostability, rapid maturation, an extended lifetime, and a monomeric nature. Additionally, mBeRFP can be paired with the Alexa 647 dye as a FRET donor to detect caspase 3 activity. This FRET pair has an extremely dynamic range and a large Förster radius (approximately 6.5 nm). To demonstrate the applicability of mBeRFP for imaging in living cells, we performed dual-color imaging of mBeRFP and CFP simultaneously excited by a single excitation source, and we demonstrated that these fluorescent proteins allow the clear visualization of the dynamics of Bax during cancer cell apoptosis. Thus, mBeRFP appears to be particularly useful for cellular imaging applications.
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.forename: Liang
authors.initials: L
authors.lastname: Wang
authors.forename: Fei
authors.initials: F
authors.lastname: Yang
authors.forename: Haiming
authors.initials: H
authors.lastname: Luo
authors.forename: Lingling
authors.initials: L
authors.lastname: Xu
authors.forename: Jinling
authors.initials: J
authors.lastname: Lu
authors.forename: Shaoqun
authors.initials: S
authors.lastname: Zeng
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D019943:Amino Acid Substitution; D017209:Apoptosis; D031541:Fluorescence Resonance Energy Transfer; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D018613:Microscopy, Confocal; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein