Johnsson K

References (7)

Title : A general method for the development of multicolor biosensors with large dynamic ranges - Hellweg_2023_Nat.Chem.Biol__
Author(s) : Hellweg L , Edenhofer A , Barck L , Huppertz MC , Frei MS , Tarnawski M , Bergner A , Koch B , Johnsson K , Hiblot J
Ref : Nat Chemical Biology , : , 2023
Abstract : Fluorescent biosensors enable the study of cell physiology with spatiotemporal resolution; yet, most biosensors suffer from relatively low dynamic ranges. Here, we introduce a family of designed Forster resonance energy transfer (FRET) pairs with near-quantitative FRET efficiencies based on the reversible interaction of fluorescent proteins with a fluorescently labeled HaloTag. These FRET pairs enabled the straightforward design of biosensors for calcium, ATP and NAD(+) with unprecedented dynamic ranges. The color of each of these biosensors can be readily tuned by changing either the fluorescent protein or the synthetic fluorophore, which enables simultaneous monitoring of free NAD(+) in different subcellular compartments following genotoxic stress. Minimal modifications of these biosensors furthermore allow their readout to be switched to fluorescence intensity, fluorescence lifetime or bioluminescence. These FRET pairs thus establish a new concept for the development of highly sensitive and tunable biosensors.
ESTHER : Hellweg_2023_Nat.Chem.Biol__
PubMedSearch : Hellweg_2023_Nat.Chem.Biol__
PubMedID: 37291200
Gene_locus related to this paper: rhoso-halo1

Title : Exchangeable HaloTag Ligands for Super-Resolution Fluorescence Microscopy - Kompa_2023_J.Am.Chem.Soc_145_3075
Author(s) : Kompa J , Bruins J , Glogger M , Wilhelm J , Frei MS , Tarnawski M , x , Este E , Heilemann M , Hiblot J , Johnsson K
Ref : Journal of the American Chemical Society , 145 :3075 , 2023
Abstract : The specific and covalent labeling of the protein HaloTag with fluorescent probes in living cells makes it a powerful tool for bioimaging. However, the irreversible attachment of the probe to HaloTag precludes imaging applications that require transient binding of the probe and comes with the risk of irreversible photobleaching. Here, we introduce exchangeable ligands for fluorescence labeling of HaloTag (xHTLs) that reversibly bind to HaloTag and that can be coupled to rhodamines of different colors. In stimulated emission depletion (STED) microscopy, probe exchange of xHTLs allows imaging with reduced photobleaching as compared to covalent HaloTag labeling. Transient binding of fluorogenic xHTLs to HaloTag fusion proteins enables points accumulation for imaging in nanoscale topography (PAINT) and MINFLUX microscopy. We furthermore introduce pairs of xHTLs and HaloTag mutants for dual-color PAINT and STED microscopy. xHTLs thus open up new possibilities in imaging across microscopy platforms for a widely used labeling approach.
ESTHER : Kompa_2023_J.Am.Chem.Soc_145_3075
PubMedSearch : Kompa_2023_J.Am.Chem.Soc_145_3075
PubMedID: 36716211
Gene_locus related to this paper: rhoso-halo1

Title : Engineered HaloTag variants for fluorescence lifetime multiplexing - Frei_2022_Nat.Methods_19_65
Author(s) : Frei MS , Tarnawski M , Roberti MJ , Koch B , Hiblot J , Johnsson K
Ref : Nat Methods , 19 :65 , 2022
Abstract : Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce HaloTag variants with either increased or decreased brightness and fluorescence lifetime compared with HaloTag7 when labeled with rhodamines. Combining these HaloTag variants enabled live-cell fluorescence lifetime multiplexing of three cellular targets in one spectral channel using a single fluorophore and the generation of a fluorescence lifetime-based biosensor. Additionally, the brightest HaloTag variant showed up to 40% higher brightness in live-cell imaging applications.
ESTHER : Frei_2022_Nat.Methods_19_65
PubMedSearch : Frei_2022_Nat.Methods_19_65
PubMedID: 34916672
Gene_locus related to this paper: rhoso-halo1

Title : Kinetic and Structural Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag - Wilhelm_2021_Biochemistry_60_2560
Author(s) : Wilhelm J , Kuhn S , Tarnawski M , Gotthard G , Tunnermann J , Tanzer T , Karpenko J , Mertes N , Xue L , Uhrig U , Reinstein J , Hiblot J , Johnsson K
Ref : Biochemistry , 60 :2560 , 2021
Abstract : The self-labeling protein tags (SLPs) HaloTag7, SNAP-tag, and CLIP-tag allow the covalent labeling of fusion proteins with synthetic molecules for applications in bioimaging and biotechnology. To guide the selection of an SLP-substrate pair and provide guidelines for the design of substrates, we report a systematic and comparative study of the labeling kinetics and substrate specificities of HaloTag7, SNAP-tag, and CLIP-tag. HaloTag7 reaches almost diffusion-limited labeling rate constants with certain rhodamine substrates, which are more than 2 orders of magnitude higher than those of SNAP-tag for the corresponding substrates. SNAP-tag labeling rate constants, however, are less affected by the structure of the label than those of HaloTag7, which vary over 6 orders of magnitude for commonly employed substrates. Determining the crystal structures of HaloTag7 and SNAP-tag labeled with fluorescent substrates allowed us to rationalize their substrate preferences. We also demonstrate how these insights can be exploited to design substrates with improved labeling kinetics.
ESTHER : Wilhelm_2021_Biochemistry_60_2560
PubMedSearch : Wilhelm_2021_Biochemistry_60_2560
PubMedID: 34339177
Gene_locus related to this paper: rhoso-halo1

Title : Luciferases with Tunable Emission Wavelengths - Hiblot_2017_Angew.Chem.Int.Ed.Engl_56_14556
Author(s) : Hiblot J , Yu Q , Sabbadini MDB , Reymond L , Xue L , Schena A , Sallin O , Hill N , Griss R , Johnsson K
Ref : Angew Chem Int Ed Engl , 56 :14556 , 2017
Abstract : We introduce luciferases whose emission maxima can be tuned to different wavelengths by chemical labeling. The luciferases are chimeras of NanoLuc with either SNAP-tag or HaloTag7. Labeling of the self-labeling tag with a fluorophore shifts the emission maximum of NanoLuc to that of the fluorophore. Luciferases with tunable colors have applications as reporter genes, for the construction of biosensors and in bioimaging.
ESTHER : Hiblot_2017_Angew.Chem.Int.Ed.Engl_56_14556
PubMedSearch : Hiblot_2017_Angew.Chem.Int.Ed.Engl_56_14556
PubMedID: 28941028

Title : Sensing acetylcholine and anticholinesterase compounds - Schena_2014_Angew.Chem.Int.Ed.Engl_53_1302
Author(s) : Schena A , Johnsson K
Ref : Angew Chem Int Ed Engl , 53 :1302 , 2014
Abstract : Acetylcholine is a key neurotransmitter, and anticholinesterase agents are essential compounds used as medical drugs, pesticides, and chemical warfare agents. A semisynthetic fluorescence-based probe for the direct, real-time detection of acetylcholine and anticholinesterase compounds is introduced. The probe possesses good sensitivity, tunable detection range, and can be selectively targeted to cell surfaces, thereby making it an attractive tool for applications in analytical chemistry and quantitative biology.
ESTHER : Schena_2014_Angew.Chem.Int.Ed.Engl_53_1302
PubMedSearch : Schena_2014_Angew.Chem.Int.Ed.Engl_53_1302
PubMedID: 24339043

Title : A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins - Lukinavicius_2013_Nat.Chem_5_132
Author(s) : Lukinavicius G , Umezawa K , Olivier N , Honigmann A , Yang G , Plass T , Mueller V , Reymond L , Correa IR, Jr. , Luo ZG , Schultz C , Lemke EA , Heppenstall P , Eggeling C , Manley S , Johnsson K
Ref : Nat Chem , 5 :132 , 2013
Abstract : The ideal fluorescent probe for bioimaging is bright, absorbs at long wavelengths and can be implemented flexibly in living cells and in vivo. However, the design of synthetic fluorophores that combine all of these properties has proved to be extremely difficult. Here, we introduce a biocompatible near-infrared silicon-rhodamine probe that can be coupled specifically to proteins using different labelling techniques. Importantly, its high permeability and fluorogenic character permit the imaging of proteins in living cells and tissues, and its brightness and photostability make it ideally suited for live-cell super-resolution microscopy. The excellent spectroscopic properties of the probe combined with its ease of use in live-cell applications make it a powerful new tool for bioimaging.
ESTHER : Lukinavicius_2013_Nat.Chem_5_132
PubMedSearch : Lukinavicius_2013_Nat.Chem_5_132
PubMedID: 23344448