Deuteration of terminal alkynes realizes simultaneous live cell Raman imaging of similar alkyne-tagged biomolecules

Egoshi, Syusuke; Dodo, Kosuke; Ohgane, Kenji; Sodeoka, Mikiko

doi:10.1039/d1ob01479j

Cited by 21 publications

(15 citation statements)

References 31 publications

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“…With similar targeted reactions, triple-bond Raman probes with the peak-shift principles have also been rationally designed and developed for sensing pH, fluoride, and metal ions . Recently, the isotope exchange reactions (especially the H/D exchange) have been harnessed on the terminal alkynes for both two-color imaging and cellular environmental sensing applications. , The H/D exchange reaction results in the dramatic alkyne Raman peak shift (>130 cm –1 ) due to both the large mass difference between D and H and the quantum coupling between the alkyne and the adjacent CD. In addition to chemical reactions, Raman peak frequency can also be tuned by the surrounding physical environment.…”

Section: Functional Raman Imaging Probesmentioning

confidence: 99%

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Wang

2022

ACS Chem. Biol.

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As an emerging optical imaging modality, stimulated Raman scattering (SRS) microscopy provides invaluable opportunities for chemical biology studies using its rich chemical information. Through rapid progress over the past decade, the development of Raman probes harnessing the chemical biology toolbox has proven to play a key role in advancing SRS microscopy and expanding biological applications. In this perspective, we first discuss the development of biorthogonal SRS imaging using small tagging of triple bonds or isotopes and highlight their unique advantages for metabolic pathway analysis and microbiology investigations. Potential opportunities for chemical biology studies integrating small tagging with SRS imaging are also proposed. We next summarize the current designs of highly sensitive and super-multiplexed SRS probes, as well as provide future directions and considerations for next-generation functional probe design. These rationally designed SRS probes are envisioned to bridge the gap between SRS microscopy and chemical biology research and should benefit their mutual development.

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Section: Functional Raman Imaging Probesmentioning

confidence: 99%

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Wang

2022

ACS Chem. Biol.

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“…This extended peak change is a result of the combined effect of adding more mass to the alkyne “oscillator” (similar to introducing heavier-mass 13 C labeling) and the coupling between the CC and adjoining C–D bonds. Such significant shifts are consistent with previous reports of deuterated acetylide compounds. , We also performed density functional theory calculations on EdU after alkyne deuteration, in which the peak shift was predicted to be 149 cm –1 , close to our experimental data. Similar peak shift values were detected for other alkyne-tagged molecules, including the cytidine analogue ethynyl-2′-deoxycytidine (EdC, 128 cm –1 shift) and the uridine analogue 5-ethynyl-uridine (EU, 134 cm –1 shift) (Figure S1a,b).…”

Section: Resultsmentioning

confidence: 99%

Alkyne-Tagged Raman Probes for Local Environmental Sensing by Hydrogen–Deuterium Exchange

Miao

2022

J. Am. Chem. Soc.

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Alkyne-tagged Raman probes have shown high promise for noninvasive and sensitive visualization of small biomolecules to understand their functional roles in live cells. However, the potential for alkynes to sense cellular environments that goes beyond imaging remains to be further explored. Here, we report a general strategy for Raman imaging-based local environment sensing by hydrogen–deuterium exchange (HDX) of terminal alkynes (termed alkyne-HDX). We first demonstrate, in multiple Raman probes, that deuterations of the alkynyl hydrogens lead to remarkable shifts of alkyne Raman peaks for about 130 cm–1, providing resolvable signals suited for imaging-based analysis with high specificity. Both our analytical derivation and experimental characterizations subsequently establish that HDX kinetics are linearly proportional to both alkyne pK as and environmental pDs. After validating the quantitative nature of this strategy, we apply alkyne-HDX to sensing local chemical and cellular environments. We establish that alkyne-HDX exhibits high sensitivity to various DNA structures and demonstrates the capacity to detect DNA structural changes in situ from UV-induced damage. We further show that this strategy is also applicable to resolve subtle pD variations in live cells. Altogether, our work lays the foundation for utilizing alkyne-HDX strategy to quantitatively sense the local environments for a broad spectrum of applications in complex biological systems.

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“…The different behaviors of saturated and unsaturated fatty acids have also been visualized using D-labeled 17-octadecynoic acid and 17-Yne oleic acid. 95 Raman imaging of various alkyne-tagged biomolecules has been reported (Figure 6). For example, SRS imaging of various alkyne-tagged metabolic precursors, EdU, EU, L-homopropargylglycine, propargyl choline, and 17-octadecynoic acid, has been used to visualize de novo synthesis of DNA, RNA, proteins, phospholipids, and triglycerides in live cells and C. elegans 96 as well as rat tissue.…”

Section: Raman Tagsmentioning

confidence: 99%

Raman Spectroscopy for Chemical Biology Research

2022

Self Cite

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In chemical biology research, various fluorescent probes have been developed and used to visualize target proteins or molecules in living cells and tissues, yet there are limitations to this technology, such as the limited number of colors that can be detected simultaneously. Recently, Raman spectroscopy has been applied in chemical biology to overcome such limitations. Raman spectroscopy detects the molecular vibrations reflecting the structures and chemical conditions of molecules in a sample and was originally used to directly visualize the chemical responses of endogenous molecules. However, our initial research to develop “Raman tags” opens a new avenue for the application of Raman spectroscopy in chemical biology. In this Perspective, we first introduce the label-free Raman imaging of biomolecules, illustrating the biological applications of Raman spectroscopy. Next, we highlight the application of Raman imaging of small molecules using Raman tags for chemical biology research. Finally, we discuss the development and potential of Raman probes, which represent the next-generation probes in chemical biology.

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Deuteration of terminal alkynes realizes simultaneous live cell Raman imaging of similar alkyne-tagged biomolecules

Abstract: Two-color Raman imaging of D-alkynes and H-alkynes makes it possible to distinguish and observe similar small molecules in live cells.

Cited by 21 publications

References 31 publications

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Alkyne-Tagged Raman Probes for Local Environmental Sensing by Hydrogen–Deuterium Exchange

Raman Spectroscopy for Chemical Biology Research

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