A Bioorthogonally Applicable, Fluorogenic, Large Stokes-Shift Probe for Intracellular Super-Resolution Imaging of Proteins.

Biomolecules
Evelin NémethPéter Kele

Abstract

Herein, we present the synthesis and application of a fluorogenic, large Stokes-shift (>100 nm), bioorthogonally conjugatable, membrane-permeable tetrazine probe, which can be excited at common laser line 488 nm and detected at around 600 nm. The applied design enabled improved fluorogenicity in the orange/red emission range, thus efficient suppression of background and autofluorescence upon imaging biological samples. Moreover, unlike our previous advanced probes, it does not require the presence of special target platforms or microenvironments to achieve similar fluorogenicity and can be generally applied, e.g., on translationally bioorthogonalized proteins. Live-cell labeling schemes revealed that the fluorogenic probe is suitable for specific labeling of intracellular proteins, site-specifically modified with a cyclooctynylated, non-canonical amino acid, even under no-wash conditions. Furthermore, the probe was found to be applicable in stimulated emission depletion (STED) super-resolution microscopy imaging using a 660 nm depletion laser. Probably the most salient feature of this new probe is that the large Stokes-shift allows dual-color labeling schemes of cellular structures using distinct excitation and the same detecti...Continue Reading

References

Jul 23, 2008·Angewandte Chemie·Mike HeilemannMarkus Sauer
Feb 4, 2010·Chemistry, an Asian Journal·Krisztina NagyPéter Kele
Jan 22, 2011·Analytical Chemistry·Knut Rurack, Monika Spieles
Jun 15, 2012·Angewandte Chemie·Jun YangNeal K Devaraj
Mar 29, 2013·Organic & Biomolecular Chemistry·András HernerPéter Kele
Apr 9, 2013·Chemical Society Reviews·Astrid-Caroline Knall, Christian Slugovc
May 29, 2013·Angewandte Chemie·Jonathan C T CarlsonRalph Weissleder
Feb 22, 2014·Annual Review of Biochemistry·Jason W Chin
Mar 25, 2014·Chemical Reviews·Kathrin Lang, Jason W Chin
Jun 12, 2014·Angewandte Chemie·Labros G MeimetisRalph Weissleder
Apr 11, 2015·Chemical Society Reviews·Gong ZhangPeng R Chen
Apr 23, 2015·Journal of the American Chemical Society·Peyton ShiehCarolyn R Bertozzi
Jun 20, 2015·Angewandte Chemie·Stefan W Hell
Jul 17, 2015·Chemistry : a European Journal·Jan-Erik HoffmannCarsten Schultz
Mar 11, 2016·Molecular BioSystems·Jonathan LotzeAnnette G Beck-Sickinger
Mar 13, 2016·Amino Acids·Christoph FreidelKalina Peneva
Jul 13, 2017·International Journal of Molecular Sciences·Chenge LiArnaud Gautier
Sep 7, 2017·Nature Reviews. Molecular Cell Biology·Steffen J SahlStefan Jakobs
Sep 8, 2015·Methods and Applications in Fluorescence·Gergely B CserépPéter Kele
Apr 21, 2018·Chemistry : a European Journal·Attila KormosPéter Kele
Dec 13, 2018·Organic & Biomolecular Chemistry·Eszter Kozma, Péter Kele
Dec 15, 2018·Journal of the American Chemical Society·Lu WangKai Johnsson

❮ Previous
Next ❯

Citations

Apr 28, 2020·Chemistry : a European Journal·Juraj GaletaMilan Vrabel
Jan 12, 2021·Methods and Applications in Fluorescence·György TörökKrisztina Németh
Jun 8, 2021·Angewandte Chemie·Vincent RigolotCedric Lion
Aug 8, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Maryana HandulaYann Seimbille
Aug 14, 2020·Journal of the American Chemical Society·Márton BojtárPéter Kele

❮ Previous
Next ❯

Methods Mentioned

BETA
fluorescence imaging
column chromatography
transfection
confocal microscopy

Software Mentioned

HELIOS
Leica Application Suite X
ImageJ
Huygens
Origin Pro

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.