Enzyme self-label-bound ATTO700 in single-molecule and super-resolution microscopy

Trumpp, Michael; Oliveras, Anna; Gonschior, Hannes; Ast, Julia; Hodson, David J.; Knaus, Petra; Lehmann, Martin; Birol, Melissa; Broichhagen, Johannes

doi:10.1039/d2cc04823j

Cited by 4 publications

(3 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The emergence of autofluorescent proteins and protein chimeras with spectroscopic characteristics suitable for FRET applications has opened the way to implement Förster resonance energy transfer in the light microscope. The study of four-dimensional (x,y,z,t) molecular-scale phenomena in live cells [ 107 ] is thus a reality that now projects beyond the optical diffraction barrier and into the realm of super-resolution light microscopy [ 108 , 109 , 110 , 111 , 112 , 113 , 114 ]. Furthermore, the ability to genetically manipulate the chemistry of donor and acceptor fluorescent proteins has expanded the spectral coverage of fluorescence microscopy in live-cell interrogation [ 110 ].…”

Section: Concluding Remarks and Future Prospectsmentioning

confidence: 99%

Carlos Gutiérrez-Merino: Synergy of Theory and Experimentation in Biological Membrane Research

Antollini,

Barrantes

2024

Molecules

View full text Add to dashboard Cite

Professor Carlos Gutiérrez-Merino, a prominent scientist working in the complex realm of biological membranes, has made significant theoretical and experimental contributions to the field. Contemporaneous with the development of the fluid-mosaic model of Singer and Nicolson, the Förster resonance energy transfer (FRET) approach has become an invaluable tool for studying molecular interactions in membranes, providing structural insights on a scale of 1–10 nm and remaining important alongside evolving perspectives on membrane structures. In the last few decades, Gutiérrez-Merino’s work has covered multiple facets in the field of FRET, with his contributions producing significant advances in quantitative membrane biology. His more recent experimental work expanded the ground concepts of FRET to high-resolution cell imaging. Commencing in the late 1980s, a series of collaborations between Gutiérrez-Merino and the authors involved research visits and joint investigations focused on the nicotinic acetylcholine receptor and its relation to membrane lipids, fostering a lasting friendship.

show abstract

Section: Concluding Remarks and Future Prospectsmentioning

confidence: 99%

Carlos Gutiérrez-Merino: Synergy of Theory and Experimentation in Biological Membrane Research

Antollini,

Barrantes

2024

Molecules

View full text Add to dashboard Cite

show abstract

“…In the far-red region of the spectrum, oxazine HaloTag ligands include ATTO655, used in living cells for single molecule localization microscopy (SMLM) of the actin cytoskeleton (Figure 2c), [15] and ATTO700 which shows good performance for STED microscopy. [16] Cyanines, with their large extinction coefficients and high photostability, have been the preferred far-red fluorophores, particularly useful for imaging deep in tissue. For example, IR800 with an extended HaloTag ligand could be applied to labelling tumor receptors in mice (Figure 2d).…”

Section: Fluorophore Ligands Based On Traditional Dye Scaffoldsmentioning

confidence: 99%

“…Across the visible range, rhodamine derivatives have been by far the most widely used fluorophores, and the next section will discuss in detail the diversity of rhodamines applied as HTLs (section 2.2). In the far‐red region of the spectrum, oxazine HaloTag ligands include ATTO655, used in living cells for single molecule localization microscopy (SMLM) of the actin cytoskeleton (Figure 2c), [15] and ATTO700 which shows good performance for STED microscopy [16] . Cyanines, with their large extinction coefficients and high photostability, have been the preferred far‐red fluorophores, particularly useful for imaging deep in tissue.…”

Section: Fluorescent and Fluorogenic Ligands For Halotag Protein Labe...mentioning

confidence: 99%

HaloTag‐Based Reporters for Fluorescence Imaging and Biosensing

2023

View full text Add to dashboard Cite

Visualizing the structure and dynamics of biomolecules is critical to understand biological function, and requires methods to fluorescently label targets of interest in their cellular context. Self‐labelling proteins, which combine a genetically encoded tag with a small‐molecule fluorophore, have attracted considerable attention for this purpose, as they can overcome limitations of fluorescent proteins. Among them, the HaloTag protein is the most broadly used, showing fast specific labelling with a small, easy to functionalize and cell‐permeant ligand. Synthetic chemistry and protein engineering have provided a portfolio of powerful imaging tools exploiting HaloTag, along with general methods to optimize and adapt them to specific applications. Here, we provide an overview of fluorescent reporters based on the HaloTag protein for imaging and biosensing, highlighting engineering strategies and general applications.

show abstract

A Bright Surprise: Live‐Cell Labeling with Negatively Charged Fluorescent Probes based on Disulfonated Rhodamines and HaloTag

Kim¹,

Stoldt

Weber³

et al. 2023

Chemistry Methods

View full text Add to dashboard Cite

Disulfonated rhodamines are photostable and bright dyes widely used in life science and optical microscopy. However, di‐sulfonated dyes were considered cell impermeable and not applicable in living cells. We challenged this assumption with 5 most popular rhodamines (Rho) having two carboxylic acid residues, versatile sulfonation patterns and emitting green (AS488), yellow (Rho530), orange (Rho565) and red (Rho590 and STAR RED) light. The probes comprising one rhodamine entity and a HaloTagTM amine (O2) ligand (x) were prepared and applied for labeling of living, Vimentin‐Halo (VIM‐Halo) expressing U‐2 OS cells. Surprisingly, we observed specific and bright staining with simplest compounds Rho590‐x, Rho565‐x and Rho530‐x bearing two negative charges; they performed well also in stimulated emission depletion (STED) microscopy. Specific staining and red shifts in absorption and emission bands were observed with other probes having one negative charge; they were prepared by native chemical ligation and esterification.

show abstract

Enzyme self-label-bound ATTO700 in single-molecule and super-resolution microscopy

Abstract: Herein, we evaluate near infrared ATTO700 as an acceptor in SNAP- and Halo-tag protein labelling for Förster Resonance Energy Transfer (FRET) by ensemble and single molecule measurements. Microscopy of cell...

Cited by 4 publications

References 17 publications

Carlos Gutiérrez-Merino: Synergy of Theory and Experimentation in Biological Membrane Research

Carlos Gutiérrez-Merino: Synergy of Theory and Experimentation in Biological Membrane Research

HaloTag‐Based Reporters for Fluorescence Imaging and Biosensing

A Bright Surprise: Live‐Cell Labeling with Negatively Charged Fluorescent Probes based on Disulfonated Rhodamines and HaloTag

Contact Info

Product

Resources

About