An Activity-Based Sensing Approach to Multiplex Mapping of Labile Copper Pools by Stimulated Raman Scattering.

Autor:	Jiang Y; Department of Chemistry, University of California, Berkeley, California 94720, United States.; Department of Chemistry, University of Rochester, Rochester, New York 14627, United States., El Khoury E; Department of Chemistry, Columbia University, New York, New York 10027, United States., Pezacki AT; Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.; Department of Chemistry, University of California, Berkeley, California 94720, United States., Qian N; Department of Chemistry, Columbia University, New York, New York 10027, United States., Oi M; Department of Chemistry, University of California, Berkeley, California 94720, United States., Torrente L; Department of Metabolism and Physiology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida 33612, United States., Miller SG; Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, Oregon 97239, United States., Ralle M; Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, Oregon 97239, United States., DeNicola GM; Department of Metabolism and Physiology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida 33612, United States., Min W; Department of Chemistry, Columbia University, New York, New York 10027, United States., Chang CJ; Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.; Department of Chemistry, University of California, Berkeley, California 94720, United States.; Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, United States.; Helen Wills Neuroscience Institute, University of California, Berkeley, California 94720, United States.
Jazyk:	angličtina
Zdroj:	Journal of the American Chemical Society [J Am Chem Soc] 2024 Dec 11; Vol. 146 (49), pp. 33324-33337. Date of Electronic Publication: 2024 Nov 25.
DOI:	10.1021/jacs.4c06296
Abstrakt:	Molecular imaging with analyte-responsive probes offers a powerful chemical approach to studying biological processes. Many reagents for bioimaging employ a fluorescence readout, but the relatively broad emission bands of this modality and the need to alter the chemical structure of the fluorophore for different signal colors can potentially limit multiplex imaging. Here, we report a generalizable approach to multiplex analyte imaging by leveraging the comparably narrow spectral signatures of stimulated Raman scattering (SRS) in activity-based sensing (ABS) mode. We illustrate this concept with two copper Raman probes (CRPs), CRP2181 and CRP2153.2 , that react selectively with loosely bound Cu(I/II) and Cu(II) ions, respectively, termed the labile copper pool, through copper-directed acyl imidazole (CDAI) chemistry. These reagents label proximal proteins in a copper-dependent manner using a dye scaffold bearing a 13 C≡N or 13 C≡ 15 N isotopic SRS tag with nearly identical physiochemical properties in terms of shape and size. SRS imaging with the CRP reagents enables duplex monitoring of changes in intracellular labile Cu(I) and Cu(II) pools upon exogenous copper supplementation or copper depletion or genetic perturbations to copper transport proteins. Moreover, CRP imaging reveals reciprocal increases in labile Cu(II) pools upon decreases in activity of the antioxidant response nuclear factor-erythroid 2-related factor 2 (NRF2) in cellular models of lung adenocarcinoma. By showcasing the use of narrow-bandwidth ABS probes for multiplex imaging of copper pools in different oxidation states and identifying alterations in labile metal nutrient pools in cancer, this work establishes a foundation for broader SRS applications in analyte-responsive imaging in biological systems.
Databáze:	MEDLINE
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