Copper‐Catalyzed Azide–Alkyne Click Chemistry for Bioconjugation

Presolski, Stanislav I.; Hong, Vu; Finn, M. G.

doi:10.1002/9780470559277.ch110148

Cited by 347 publications

(306 citation statements)

References 51 publications

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“…The slides were then cured for 2 min at 100°C before a second spin coating at 2000 rpm for 20 sec with a solution in toluene with the following: 0.5% polystyrene (Sigma, 182427), 0.005% azide-terminated polystyrene (Sigma, 699772), and M280 streptavidin beads (ThermoFisher, 11205D) (2 µL of beads for 200 µL of polystyrene solution prerinsed with methanol). After assembling into an imaging chamber with the spin-coated sides facing inward, biotin was conjugated by click chemistry as reported (Presolski et al 2011) with the following key specific reagents: biotin-PEG5000-C12-Alkyne (Baseclick) and THPTA-ligand (Baseclick). The chambers were rinsed with standard PBS buffer supplemented with 0.1% Tween 20 before use.…”

Section: In Vitro Single-molecule Imaging and Data Analysismentioning

confidence: 99%

Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis

et al. 2016

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Transcription of protein-encoding genes in eukaryotic cells requires the coordinated action of multiple general transcription factors (GTFs) and RNA polymerase II (Pol II). A "step-wise" preinitiation complex (PIC) assembly model has been suggested based on conventional ensemble biochemical measurements, in which protein factors bind stably to the promoter DNA sequentially to build a functional PIC. However, recent dynamic measurements in live cells suggest that transcription factors mostly interact with chromatin DNA rather transiently. To gain a clearer dynamic picture of PIC assembly, we established an integrated in vitro single-molecule transcription platform reconstituted from highly purified human transcription factors and complemented it by live-cell imaging. Here we performed real-time measurements of the hierarchal promoter-specific binding of TFIID, TFIIA, and TFIIB. Surprisingly, we found that while promoter binding of TFIID and TFIIA is stable, promoter binding by TFIIB is highly transient and dynamic (with an average residence time of 1.5 sec). Stable TFIIB-promoter association and progression beyond this apparent PIC assembly checkpoint control occurs only in the presence of Pol II-TFIIF. This transient-to-stable transition of TFIIB-binding dynamics has gone undetected previously and underscores the advantages of single-molecule assays for revealing the dynamic nature of complex biological reactions.

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Section: In Vitro Single-molecule Imaging and Data Analysismentioning

confidence: 99%

Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis

et al. 2016

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“…The reaction was carried out in 500 mL TEAA buffer and le in darkness for 1 h under sonication to yield compound 3. 21,22 Compound 3 was then puried using a C18 column and the reaction products were monitored at 200 nm and 280 nm. 23 The produced Nd-DOTA-KE 3 was then used for SA labelling 4.…”

Section: Preparation Of Ln-dota-kementioning

confidence: 99%

Detection of sulfenic acid in intact proteins by mass spectrometric techniques: application to serum samples

et al. 2017

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Formation of cysteine sulfenic acid (SA) is considered a transient state of thiol oxidation in living organisms that can be either reduced back or continue to result in the formation of sulfinic and sulfonic acids. As any disturbance in oxidation is correlated to age-related diseases such as cancer and Alzheimer's disease, the detection of SA transient state formed a sensor for such redox-mediated events. Thus, detection of low amounts of SA is critical in order to prevent further oxidative damage of cells and tissues and for this aim specific strategies have to be developed. In this work, detection and quantification of induced SA in human serum albumin is reported by specific capture using alkyne bketoester (KE) previously linked to a lanthanide (Ln)-containing chelator (Ln-DOTA, where DOTA is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid). SA formation was induced by hydrogen peroxide to mimic oxidative conditions produced in living cells by ROS and was detected using molecular and elemental mass spectrometric (MS) techniques. The developed strategy has been further applied to the determination of SA-induced formation in human serum by using affinity chromatography for purification of albumin followed by inductively coupled plasma mass spectrometry (ICP-MS) to monitor the formed SA linked to Ln-DOTA-KE in combination with isotope dilution analysis (IDA) for absolute quantification. Quantitative results showed levels of oxidative damage regarding SA formation in up to 40% of the albumin present.

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“…In general, molecular surface modifications of these particles can be performed by grafting methods [8], whereas biomedically useful surface modifications are mostly achieved via cycloaddition reactions, e.g., the famous click reaction [9][10][11], biotin-streptavidin interactions [12], and carbodiimide coupling reactions [13]. By these techniques, immobilization of antibodies, drugs, and vitamin units is achieved and widely reported in the literature [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Asymmetric attachment and functionalization of plasmonic nanoparticles on ceramic interfaces

et al. 2018

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The demands for materials that integrate more than one functional imaging or therapeutic unit are of increasing interest for biomedical applications. Here, we present the step-by-step preparation of asymmetric and optically active particles, namely, Gd 2 O 3 @Ag, Gd 2 O 3 @Au, SiO 2 -N 3 @Au, and SiO 2 -SH@Au . Successful attachment of plasmonic nanoparticles to the surface of metal-oxide spheres without necessity of a potentially toxic inter-adhesive layer was proven by optical methods as well as X-ray photoelectron spectroscopy. The combination of optical and magnetic properties as present in Gd 2 O 3 @ Ag and Gd 2 O 3 @Au Janus-type particles leads to dual-imaging probes for optical and magnetic resonance imaging. In addition, functional groups, such as azide groups, were linked to the surface of silica particles previous to Au nanoparticle attachment. Subsequent site-selective click reactions with 5-FAM were successfully performed as demonstrated by UV-Vis measurements. All described systems exhibited excellent long-term stability and can, therefore, be considered as promising candidates for theranostic applications. Graphical abstract

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Copper‐Catalyzed Azide–Alkyne Click Chemistry for Bioconjugation

Cited by 347 publications

References 51 publications

Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis

Rapid dynamics of general transcription factor TFIIB binding during preinitiation complex assembly revealed by single-molecule analysis

Detection of sulfenic acid in intact proteins by mass spectrometric techniques: application to serum samples

Asymmetric attachment and functionalization of plasmonic nanoparticles on ceramic interfaces

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