Click-free imaging of carbohydrate trafficking in live cells using an azido photothermal probe.

Autor:	Xia Q; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, United States.; Photonics Center, Boston University, Boston, Massachusetts 02215, United States., Perera HA; Department of Chemistry, University of Massachusetts, Lowell, Massachusetts 01854, United States., Bolarinho R; Department of Chemistry, Boston University, Boston, Massachusetts 02215, United States., Piskulich ZA; Department of Chemistry, Boston University, Boston, Massachusetts 02215, United States., Guo Z; Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, United States., Yin J; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, United States., He H; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, United States., Li M; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, United States., Ge X; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, United States., Cui Q; Department of Chemistry, Boston University, Boston, Massachusetts 02215, United States., Ramström O; Department of Chemistry, University of Massachusetts, Lowell, Massachusetts 01854, United States.; Department of Chemistry and Biomedical Sciences, Linnaeus University, SE-39182 Kalmar, Sweden., Yan M; Department of Chemistry, University of Massachusetts, Lowell, Massachusetts 01854, United States., Cheng JX; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, United States.; Photonics Center, Boston University, Boston, Massachusetts 02215, United States.; Department of Chemistry, Boston University, Boston, Massachusetts 02215, United States.; Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, United States.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Mar 12. Date of Electronic Publication: 2024 Mar 12.
DOI:	10.1101/2024.03.08.584185
Abstrakt:	Real-time tracking of intracellular carbohydrates remains challenging. While click chemistry allows bio-orthogonal tagging with fluorescent probes, the reaction permanently alters the target molecule and only allows a single snapshot. Here, we demonstrate click-free mid-infrared photothermal (MIP) imaging of azide-tagged carbohydrates in live cells. Leveraging the micromolar detection sensitivity for 6-azido-trehalose (TreAz) and the 300-nm spatial resolution of MIP imaging, the trehalose recycling pathway in single mycobacteria, from cytoplasmic uptake to membrane localization, is directly visualized. A peak shift of azide in MIP spectrum further uncovers interactions between TreAz and intracellular protein. MIP mapping of unreacted azide after click reaction reveals click chemistry heterogeneity within a bacterium. Broader applications of azido photothermal probes to visualize the initial steps of the Leloir pathway in yeasts and the newly synthesized glycans in mammalian cells are demonstrated. Competing Interests: Competing interests. JXC declares interests with Photothermal Spectroscopy Corp., which did not support this study. Other authors declare no competing interests.
Databáze:	MEDLINE
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