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Imaging complex protein metabolism in live organisms by stimulated Raman scattering microscopy with isotope labeling

Wei, Lu and Shen, Yihui and Xu, Fang and Hu, Fanghao and Harrington, Jamie K. and Targoff, Kimara L. and Min, Wei (2015) Imaging complex protein metabolism in live organisms by stimulated Raman scattering microscopy with isotope labeling. ACS Chemical Biology, 10 (3). pp. 901-908. ISSN 1554-8929. PMCID PMC4610303.

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Protein metabolism, consisting of both synthesis and degradation, is highly complex, playing an indispensable regulatory role throughout physiological and pathological processes. Over recent decades, extensive efforts, using approaches such as autoradiography, mass spectrometry, and fluorescence microscopy, have been devoted to the study of protein metabolism. However, noninvasive and global visualization of protein metabolism has proven to be highly challenging, especially in live systems. Recently, stimulated Raman scattering (SRS) microscopy coupled with metabolic labeling of deuterated amino acids (D-AAs) was demonstrated for use in imaging newly synthesized proteins in cultured cell lines. Herein, we significantly generalize this notion to develop a comprehensive labeling and imaging platform for live visualization of complex protein metabolism, including synthesis, degradation, and pulse-chase analysis of two temporally defined populations. First, the deuterium labeling efficiency was optimized, allowing time-lapse imaging of protein synthesis dynamics within individual live cells with high spatial-temporal resolution. Second, by tracking the methyl group (CH3) distribution attributed to pre-existing proteins, this platform also enables us to map protein degradation inside live cells. Third, using two subsets of structurally and spectroscopically distinct D-AAs, we achieved two-color pulse-chase imaging, as demonstrated by observing aggregate formation of mutant hungtingtin proteins. Finally, going beyond simple cell lines, we demonstrated the imaging ability of protein synthesis in brain tissues, zebrafish, and mice in vivo. Hence, the presented labeling and imaging platform would be a valuable tool to study complex protein metabolism with high sensitivity, resolution, and biocompatibility for a broad spectrum of systems ranging from cells to model animals and possibly to humans.

Item Type:Article
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URLURL TypeDescription CentralArticle Information
Wei, Lu0000-0001-9170-2283
Hu, Fanghao0000-0002-8659-4027
Min, Wei0000-0003-2570-3557
Additional Information:© 2015 American Chemical Society. ACS AuthorChoice - This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. Received: September 30, 2014. Accepted: January 5, 2015. Published: January 5, 2015. We thank J. Jackson and C. Dupre for assistance with the brain slices and J. C. Tapia, M. C. Wang, Z. Chen, D. Peterka, and R. Yuste for helpful discussions. We are grateful to Y. Shin for technical assistance with the in vivo mice experiments. W.M. acknowledges support from Columbia University, an National Institutes of Health Director’s New Innovator Award, the U.S. Army Research Office (W911NF-12-1-0594), the Brain Research Foundation, and an Alfred P. Sloan Research Fellowship. Author Contributions: L.W., Y.S., F.X., F.H., J.K.H., and K.L.T. performed experiments and analyzed data. L.W., Y.S., and W.M. designed the experiments. L.W. and W.M. conceived the concept and wrote the article. The authors declare the following competing financial interest(s): Columbia University has filed a patent application based on this work.
Funding AgencyGrant Number
Columbia UniversityUNSPECIFIED
Army Research Office (ARO)W911NF-12-1-0594
Brain Research FoundationUNSPECIFIED
Alfred P. Sloan FoundationUNSPECIFIED
Issue or Number:3
PubMed Central ID:PMC4610303
Record Number:CaltechAUTHORS:20180608-131444540
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Official Citation:Imaging Complex Protein Metabolism in Live Organisms by Stimulated Raman Scattering Microscopy with Isotope Labeling Lu Wei, Yihui Shen, Fang Xu, Fanghao Hu, Jamie K. Harrington, Kimara L. Targoff, and Wei Min ACS Chemical Biology 2015 10 (3), 901-908 DOI: 10.1021/cb500787b
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:86926
Deposited By: George Porter
Deposited On:08 Jun 2018 23:06
Last Modified:09 Mar 2020 13:19

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