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, MA 02215, USA.; Photonics Center, Boston University, Boston, MA 02215, USA., Perera HA; Department of Chemistry, University of Massachusetts, Lowell, MA 01854, USA., Bolarinho R; Department of Chemistry, Boston University, Boston, MA 02215, USA., Piskulich ZA; Department of Chemistry, Boston University, Boston, MA 02215, USA., Guo Z; Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA., Yin J; Department of Electrical and Computer Engineering, Boston University, Boston, MA 02215, USA., He H; Department of Electrical and Computer Engineering, Boston University, Boston, MA 02215, USA., Li M; Department of Electrical and Computer Engineering, Boston University, Boston, MA 02215, USA., Ge X; Department of Electrical and Computer Engineering, Boston University, Boston, MA 02215, USA., Cui Q; Department of Chemistry, Boston University, Boston, MA 02215, USA., Ramström O; Department of Chemistry, University of Massachusetts, Lowell, MA 01854, USA.; Department of Chemistry and Biomedical Sciences, Linnaeus University, SE-39182 Kalmar, Sweden., Yan M; Department of Chemistry, University of Massachusetts, Lowell, MA 01854, USA., Cheng JX; Department of Electrical and Computer Engineering, Boston University, Boston, MA 02215, USA.; Photonics Center, Boston University, Boston, MA 02215, USA.; Department of Chemistry, Boston University, Boston, MA 02215, USA.; Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
Jazyk: angličtina
Zdroj: Science advances [Sci Adv] 2024 Aug 23; Vol. 10 (34), pp. eadq0294. Date of Electronic Publication: 2024 Aug 21.
DOI: 10.1126/sciadv.adq0294
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.
Databáze: MEDLINE