scholarly journals Site-specific near-infrared fluorescent labelling of proteins on cysteine residues withmeso-chloro-substituted heptamethine cyanine dyes

2018 ◽  
Vol 16 (45) ◽  
pp. 8831-8836 ◽  
Author(s):  
Coline Canovas ◽  
Pierre-Simon Bellaye ◽  
Mathieu Moreau ◽  
Anthony Romieu ◽  
Franck Denat ◽  
...  

meso-Chloroheptamethine cyanine dyes react selectively with cysteine residues in proteins to give site-specifically labelled NIR fluorescent bioconjugates.

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2900 ◽  
Author(s):  
Chen-Ming Lin ◽  
Syed Usama ◽  
Kevin Burgess

Convenient labeling of proteins is important for observing its function under physiological conditions. In tissues particularly, heptamethine cyanine dyes (Cy-7) are valuable because they absorb in the near-infrared (NIR) region (750–900 nm) where light penetration is maximal. In this work, we found Cy-7 dyes with a meso-Cl functionality covalently binding to proteins with free Cys residues under physiological conditions (aqueous environments, at near neutral pH, and 37 °C). It transpired that the meso-Cl of the dye was displaced by free thiols in protein, while nucleophilic side-chains from amino acids like Tyr, Lys, and Ser did not react. This finding shows a new possibility for convenient and selective labeling of proteins with NIR fluorescent probes.


1997 ◽  
Vol 62 (26) ◽  
pp. 9387-9387 ◽  
Author(s):  
Narasimhachari Narayanan ◽  
Lucjan Strekowski ◽  
Malgorzata Lipowska ◽  
Gabor Patonay

Biochemistry ◽  
1994 ◽  
Vol 33 (38) ◽  
pp. 11637-11644 ◽  
Author(s):  
Robert J. Kreitman ◽  
Raj K. Puri ◽  
Pamela Leland ◽  
Byungkook Lee ◽  
Ira Pastan

BIOPHYSICS ◽  
2021 ◽  
Vol 66 (1) ◽  
pp. 25-31
Author(s):  
A. Yu. Ikonnikova ◽  
V. E. Shershov ◽  
Yu. V. Moroz ◽  
V. A. Vasiliskov ◽  
S. A. Lapa ◽  
...  

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Marie Synakewicz ◽  
Daniela Bauer ◽  
Matthias Rief ◽  
Laura S. Itzhaki

Abstract Accurate and stable site-specific attachment of DNA molecules to proteins is a requirement for many single-molecule force spectroscopy techniques. The most commonly used method still relies on maleimide chemistry involving cysteine residues in the protein of interest. Studies have consequently often focused on model proteins that either have no cysteines or with a small number of cysteines that can be deleted so that cysteines can then be introduced at specific sites. However, many proteins, especially in eukaryotes, contain too many cysteine residues to be amenable to this strategy, and therefore there is tremendous need for new and broadly applicable approaches to site-specific conjugation. Here we present bioorthogonal approaches for making DNA-protein conjugates required in force spectroscopy experiments. Unnatural amino acids are introduced site-specifically and conjugated to DNA oligos bearing the respective modifications to undergo either strain-promoted azidealkyne cycloaddition (SPAAC) or inverse-electron-demand Diels-Alder (IE-DA) reactions. We furthermore show that SPAAC is compatible with a previously published peptide-based attachment approach. By expanding the available toolkit to tag-free methods based on bioorthogonal reactions, we hope to enable researchers to interrogate the mechanics of a much broader range of proteins than is currently possible.


Materials ◽  
2020 ◽  
Vol 13 (9) ◽  
pp. 2083
Author(s):  
Tiago D. Martins ◽  
Eurico Lima ◽  
Renato E. Boto ◽  
Diana Ferreira ◽  
José R. Fernandes ◽  
...  

Photodynamic therapy is a medical modality developed for the treatment of several diseases of oncological and non-oncological etiology that requires the presence of a photosensitizer, light and molecular oxygen, which combined will trigger physicochemical reactions responsible for reactive oxygen species production. Given the scarcity of photosensitizers that exhibit desirable characteristics for its potential application in this therapeutic strategy, the main aims of this work were the study of the photophysical and photochemical properties and the photobiological activity of several dicyanomethylene squaraine cyanine dyes. Thus, herein, the study of their aggregation character, photobleaching and singlet oxygen production ability, and the further application of the previously synthesized dyes in Caco-2 and HepG2 cancer cell lines, to evaluate their phototherapeutic effects, are described. Dicyanomethylene squaraine dyes exhibited moderate light-stability and, despite the low singlet oxygen quantum yields, were a core of dyes that exhibited relevant in vitro photodynamic activity, as there was an evident increase in the toxicity of some of the tested dyes exclusive to radiation treatments.


2015 ◽  
Vol 22 (7) ◽  
pp. 965-975 ◽  
Author(s):  
Neal S. Gould ◽  
Perry Evans ◽  
Pablo Martínez-Acedo ◽  
Stefano M. Marino ◽  
Vadim N. Gladyshev ◽  
...  

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