scholarly journals Advanced Fluorescent Polymer Probes for the Site-Specific Labeling of Proteins in Live Cells Using the HaloTag Technology

ACS Omega ◽  
2019 ◽  
Vol 4 (7) ◽  
pp. 12841-12847
Author(s):  
Thomas Berki ◽  
Anush Bakunts ◽  
Damien Duret ◽  
Laura Fabre ◽  
Catherine Ladavière ◽  
...  
Keyword(s):  
Live Cells ◽  
Site Specific ◽  
Fluorescent Polymer

scholarly journals Engineering of a zero cross-talk fluorescent polymer nanoprobe for self-referenced ratiometric imaging of lysosomal hypochlorous acid in living cells

2020 ◽  
Vol 4 (3) ◽  
pp. 862-868 ◽  
Author(s):  
Hongqing Wei ◽  
Rongjin Zeng ◽  
Shenglan Wang ◽  
Chong-Hua Zhang ◽  
Shu Chen ◽  
...  
Keyword(s):  
Cross Talk ◽  
Hypochlorous Acid ◽  
Living Cells ◽  
Live Cells ◽  
Ratiometric Imaging ◽  
Fluorescent Polymer

A zero cross-talk fluorescent polymer nanoprobe with a self-referenced ratiometric effect was rationally designed and synthesized for lysosomal hypochlorous acid (HClO) imaging in live cells.

scholarly journals An aluminium-based fluorinated counterion for enhanced encapsulation and emission of dyes in biodegradable polymer nanoparticles

2017 ◽  
Vol 1 (11) ◽  
pp. 2309-2316 ◽  
Author(s):  
Bohdan Andreiuk ◽  
Andreas Reisch ◽  
Vasyl G. Pivovarenko ◽  
Andrey S. Klymchenko
Keyword(s):  
Biodegradable Polymer ◽  
Polymer Nanoparticles ◽  
Live Cells ◽  
Cationic Dye ◽  
Fluorescent Polymer

An aluminium-based anion allows preparation of ultra-bright fluorescent polymer nanoparticles with nearly quantitative cationic dye encapsulation, minimized self-quenching and no dye leakage in live cells.

scholarly journals Selected peptide-based fluorescent probes for biological applications

2020 ◽  
Vol 16 ◽  
pp. 2971-2982
Author(s):  
Debabrata Maity
Keyword(s):  
Fluorescent Probes ◽  
Direct Detection ◽  
Aqueous Media ◽  
Live Cells ◽  
Biological Applications ◽  
Site Specific ◽  
Specific Manner ◽  
Environmentally Sensitive ◽  
Living Organisms ◽  
A Site

To understand the molecular interactions, present in living organisms and their environments, chemists are trying to create novel chemical tools. In this regard, peptide-based fluorescence techniques have attracted immense interest. Synthetic peptide-based fluorescent probes are advantageous over protein-based sensors, since they are synthetically accessible, more stable, and can be easily modified in a site-specific manner for selective biological applications. Peptide receptors labeled with environmentally sensitive/FRET fluorophores have allowed direct detection/monitoring of biomolecules in aqueous media and in live cells. In this review, key peptide-based approaches for different biological applications are presented.

scholarly journals Design of Fluorescent Probes for Bioorthogonal Labeling of Carbonylation in Live Cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hazel Erkan ◽  
Dilek Telci ◽  
Ozlem Dilek
Keyword(s):  
Cell Lines ◽  
Rapid Development ◽  
Covalent Binding ◽  
Dermal Fibroblasts ◽  
Live Cells ◽  
Human Dermal Fibroblasts ◽  
Bioorthogonal Reaction ◽  
Site Specific ◽  
Bioorthogonal Labeling ◽  
Diagnostic Approaches

Abstract With the rapid development of chemical biology, many diagnostic fluorophore-based tools were introduced to specific biomolecules by covalent binding. Bioorthogonal reactions have been widely utilized to manage challenges faced in clinical practice for early diagnosis and treatment of several tumor samples. Herein, we designed a small molecule fluorescent-based biosensor, 2Hydrazine-5nitrophenol (2Hzin5NP), which reacts with the carbonyl moiety of biomolecules through bioorthogonal reaction, therefore can be utilized for the detection of biomolecule carbonylation in various cancer cell lines. Our almost non-fluorescent chemical probe has a fast covalent binding with carbonyl moieties at neutral pH to form a stable fluorescent hydrazone product leading to a spectroscopic alteration in live cells. Microscopic and fluorometric analyses were used to distinguish the exogenous and endogenous ROS induced carbonylation profile in human dermal fibroblasts along with A498 primary site and ACHN metastatic site renal cell carcinoma (RRC) cell lines. Our results showed that carbonylation level that differs in response to exogenous and endogenous stress in healthy and cancer cells can be detected by the newly synthesized bioorthogonal fluorescent probe. Our results provide new insights into the development of novel bioorthogonal probes that can be utilized in site-specific carbonylation labeling to enhance new diagnostic approaches in cancer.

Ni–nitrilotriacetic acid-modified quantum dots as a site-specific labeling agent of histidine-tagged proteins in live cells

2008 ◽  
pp. 1910 ◽  
Author(s):  
Junwon Kim ◽  
Hye-Young Park ◽  
Jaeseung Kim ◽  
Jiyoung Ryu ◽  
Do Yoon Kwon ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Nitrilotriacetic Acid ◽  
Live Cells ◽  
Site Specific ◽  
A Site

scholarly journals Time-lapse Imaging of Individual BKCa Channels in Live Cells Using Site-specific Labeling of Quantum Dots

2009 ◽  
Vol 96 (3) ◽  
pp. 35a
Author(s):  
Ji-Yeon Kim ◽  
Haedeun Kim ◽  
Sungho Chang ◽  
Chul-Seung Park
Keyword(s):  
Quantum Dots ◽  
Time Lapse ◽  
Live Cells ◽  
Bkca Channels ◽  
Site Specific ◽  
Time Lapse Imaging

scholarly journals Site‐Specific Encoding of Photoactivity in Antibodies Enables Light‐Mediated Antibody–Antigen Binding on Live Cells

2019 ◽  
Vol 58 (50) ◽  
pp. 17986-17993 ◽  
Author(s):  
Thomas Bridge ◽  
Saher A. Shaikh ◽  
Paul Thomas ◽  
Joaquin Botta ◽  
Peter J. McCormick ◽  
...  
Keyword(s):  
Live Cells ◽  
Antigen Binding ◽  
Site Specific

scholarly journals Thiophene-derived polymer dots for imaging endocytic compartments in live cells and broad-spectrum bacterial killing

2017 ◽  
Vol 1 (1) ◽  
pp. 152-157 ◽  
Author(s):  
Kenath Priyanka Prasad ◽  
Aung Than ◽  
Nan Li ◽  
Mahasin Alam SK ◽  
Hongwei Duan ◽  
...  
Keyword(s):  
Broad Spectrum ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Live Cells ◽  
Bacterial Killing ◽  
Polymer Dots ◽  
Fluorescent Polymer ◽  
Endocytic Compartments

A new multi-functional fluorescent polymer dot was readily synthesized and used for live cell imaging and broad-spectrum bacterial killing.

scholarly journals Differential Mast Cell Outcomes Are Sensitive to FcεRI-Syk Binding Kinetics

2017 ◽  
Author(s):  
Samantha L. Schwartz ◽  
Cédric Cleyrat ◽  
Mark Olah ◽  
Peter Relich ◽  
Genevieve Phillips ◽  
...  
Keyword(s):  
Single Molecule ◽  
Protein Interactions ◽  
Calcium Release ◽  
Aggregate Size ◽  
Live Cells ◽  
Single Molecule Imaging ◽  
Site Specific ◽  
Sh2 Domains ◽  
Downstream Signaling ◽  
Transient Nature

AbstractCrosslinking of IgE-bound FcεRI triggers multiple cellular responses, including degranulation and cytokine production. Signaling is dependent on recruitment of Syk via docking of its dual SH2 domains to phosphorylated tyrosines within the FcεRI immunoreceptor tyrosine-based activation motifs. Using single molecule imaging in live cells, we directly visualized and quantified the binding of individual mNeonGreen-tagged Syk molecules as they associated with the plasma membrane after FcεRI activation. We found that Syk colocalizes transiently to FcεRI and that Syk-FcεRI binding dynamics are independent of receptor aggregate size. Substitution of glutamic acid for tyrosine between the Syk SH2 domains (SykY130E) led to an increased Syk-FcεRI off-rate, loss of site-specific Syk autophosphorylation, and impaired downstream signaling. CRISPR-Cas9 engineered cells expressing only SykY130E were deficient in antigen-stimulated calcium release, degranulation and production of some cytokines (TNF-a, IL-3) but not others (MCP-1, IL-4). We propose that kinetic discrimination along the FcεRI signaling pathway occurs at the level of Syk-FcεRI interactions, with key outcomes dependent upon sufficiently long-lived Syk binding events.SummarySchwartz et al. use single molecule imaging to quantify the transient nature of FcεRI-Syk interactions in live mast cells. A functional mutation that increases Syk off-rate leads to loss of site-specific Syk phosphorylation and impaired signaling, highlighting the importance of finely tuned protein interactions in directing cellular outcomes.

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