Fluorogenic Azidofluoresceins for Biological Imaging

Shieh, Peyton; Hangauer, Matthew J.; Bertozzi, Carolyn R.

doi:10.1021/ja308203h

Cited by 142 publications

(118 citation statements)

References 35 publications

(60 reference statements)

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“…The four analogs containing the same pendant aryl rings from our previous study (30) (1 to 4, Fig. 3) did not display significant fluorescence enhancement upon triazole formation.…”

Section: Significancementioning

confidence: 60%

“…In previous work, we used PeT from the pendant aryl ring to control the fluorescence quantum yield of fluorogenic azidofluoresceins (30). Conversion of their aryl azides to the corresponding triazoles upon Cu-catalyzed or copper-free "click" reaction with alkynes decreased their aryl ring electron density, which reduced PeT efficiency and resulted in fluorescence enhancement.…”

mentioning

confidence: 99%

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Imaging bacterial peptidoglycan with near-infrared fluorogenic azide probes

Shieh¹,

Siegrist²,

Cullen³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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191

139

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Fluorescent probes designed for activation by bioorthogonal chemistry have enabled the visualization of biomolecules in living systems. Such activatable probes with near-infrared (NIR) emission would be ideal for in vivo imaging but have proven difficult to engineer. We present the development of NIR fluorogenic azide probes based on the Si-rhodamine scaffold that undergo a fluorescence enhancement of up to 48-fold upon reaction with terminal or strained alkynes. We used the probes for mammalian cell surface imaging and, in conjunction with a new class of cyclooctyne D-amino acids, for visualization of bacterial peptidoglycan without the need to wash away unreacted probe.

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“…The four analogs containing the same pendant aryl rings from our previous study (30) (1 to 4, Fig. 3) did not display significant fluorescence enhancement upon triazole formation.…”

Section: Significancementioning

confidence: 60%

mentioning

confidence: 99%

Imaging bacterial peptidoglycan with near-infrared fluorogenic azide probes

Shieh¹,

Siegrist²,

Cullen³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

191

139

View full text Add to dashboard Cite

show abstract

“…It must be noted that the vast majority of fluorogenic probes feature an aromatic azide, which obviously will have a major impact on determination of reaction rate constant in absolute terms, as was for example found by us for a range of BCN derivatives [36]. Other more recent fluorogenic azides, as developed by Shieh et al [51] and Herner et al [26] and even a fluorogenic azide substrate for generation of a near-infrared (NIR) triazole dye [52], are depicted in Fig. 8.…”

Section: Tools To Quantify Spaac Reaction Ratesmentioning

confidence: 82%

Strain-Promoted 1,3-Dipolar Cycloaddition of Cycloalkynes and Organic Azides

Dommerholt

Rutjes

Delft

2016

Cycloadditions in Bioorthogonal Chemistry

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A nearly forgotten reaction discovered more than 60 years ago-the cycloaddition of a cyclic alkyne and an organic azide, leading to an aromatic triazole-enjoys a remarkable popularity. Originally discovered out of pure chemical curiosity, and dusted off early this century as an efficient and clean bioconjugation tool, the usefulness of cyclooctyne-azide cycloaddition is now adopted in a wide range of fields of chemical science and beyond. Its ease of operation, broad solvent compatibility, 100 % atom efficiency, and the high stability of the resulting triazole product, just to name a few aspects, have catapulted this so-called strain-promoted azide-alkyne cycloaddition (SPAAC) right into the top-shelf of the toolbox of chemical biologists, material scientists, biotechnologists, medicinal chemists, and more. In this chapter, a brief historic overview of cycloalkynes is provided first, along with the main synthetic strategies to prepare cycloalkynes and their chemical reactivities. Core aspects of the strain-promoted reaction of cycloalkynes with azides are covered, as well as tools to achieve further reaction acceleration by means of modulation of cycloalkyne structure, nature of azide, and choice of solvent.

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“…In addition to directly producing orthogonally functionalized natural product analogues that can be subjected to facile chemical modification for drug screening, our work may significantly advance the field of natural product research through coupling with diverse azido analytical handles. For example, using a fluorogenic probe 37,38 , in situ detection and quantification of natural products in producing cell cultures can be accomplished for the first time. Furthermore, since azide-alkyne click chemistry continues to serve as a powerful tool in drug discovery and chemical biology 5,39,40 , our findings open the door to numerous biological applications in which in situ enzymatic generation of a terminal alkyne is required or preferred.…”

Section: Discussionmentioning

confidence: 99%

De novo biosynthesis of terminal alkyne-labeled natural products

2014

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Abstract:The terminal alkyne is a functionality widely used in organic synthesis, pharmaceutical science, material science, and bioorthogonal chemistry. This functionality is also found in acetylenic natural products, but the underlying biosynthetic pathways for its formation are not well understood. Here we report the characterization of the first carrier proteindependent terminal alkyne biosynthetic machinery in microbes. We further demonstrate that this enzymatic machinery can be exploited for the in situ generation and incorporation of terminal alkynes into two natural product scaffolds in E. coli. These results highlight the prospect for tagging major classes of natural products, including polyketides and polyketide/non-ribosomal peptide hybrids, using biosynthetic pathway engineering.2 Natural products are important small molecules widely used as drugs, pesticides, herbicides, and biological probes. Tagging natural products with a unique chemical handle enables the visualization, enrichment, quantification, and mode of action study of natural products through bioorthogonal chemistry [1][2][3][4] . One prevalent bioorthogonal reaction is the triazole-forming azide-alkyne [3+2] cycloaddition, often referred to as "click" chemistry 5 . This reaction has enabled selective imaging and study of azide-or alkyne-labeled glycans, proteins, nucleic acids and lipids. Despite the success with macromolecules, the labeling of natural products has not been adequately explored. It is often challenging to obtain tagged natural products through total synthesis because of their structural complexity, or through semi-synthesis because of their chemical lability and the limited supply of most natural products. Alternatively, precursor-directed biosynthesis (PDB) may be employed to produce azide-or alkyne-labeled natural products based on the promiscuity of biosynthetic machinery. Mainly used as a tool to introduce structural diversity, PDB has allowed us and other researchers to generate labeled natural products 3,[6][7][8] . However, the coexistence of diffusible precursors and final products 9 with the same chemical handle introduces significant background in the PDB production system, making it incompatible with in situ bioorthogonal chemical transformations. We report here the de novo biosynthesis of alkyne-labeled natural products without the feeding of alkynoic precursors by characterizing and engineering the novel terminal alkyne synthetic machinery that living systems offer.Many acetylenic natural products contain a terminal alkyne functionality, which seems to be crucial for their bioactivity (Fig. 1) 10,11 . Terminal alkynes can be formed by acetylenases, a special family of desaturases that catalyze O 2 -dependent dehydrogenation of C-C bonds in a diiron dependent mechanism 12 . Several membrane-bound acetylenases have recently been 3 identified from plants, insects, fungi, and bacteria 4,10,[13][14][15][16][17] , but all of these reports have been limited to bioinformatics or in vivo studies through mutagenesis or ...

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Fluorogenic Azidofluoresceins for Biological Imaging

Cited by 142 publications

References 35 publications

Imaging bacterial peptidoglycan with near-infrared fluorogenic azide probes

Imaging bacterial peptidoglycan with near-infrared fluorogenic azide probes

Strain-Promoted 1,3-Dipolar Cycloaddition of Cycloalkynes and Organic Azides

De novo biosynthesis of terminal alkyne-labeled natural products

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