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Live‐Cell Quantitative Imaging of Proteome Degradation by Stimulated Raman Scattering
Author(s) -
Shen Yihui,
Xu Fang,
Wei Lu,
Hu Fanghao,
Min Wei
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201310725
Subject(s) - neurodegeneration , proteome , chemistry , protein degradation , huntingtin , biophysics , cell , microbiology and biotechnology , degradation (telecommunications) , proteomics , viability assay , biochemistry , biology , mutant , telecommunications , computer science , medicine , disease , pathology , gene
Protein degradation is a regulatory process essential to cell viability and its dysfunction is implicated in many diseases, such as aging and neurodegeneration. In this report, stimulated Raman scattering microscopy coupled with metabolic labeling with 13 C‐phenylalanine is used to visualize protein degradation in living cells with subcellular resolution. We choose the ring breathing modes of endogenous 12 C‐phenylalanine and incorporated 13 C‐phenylalanine as protein markers for the original and nascent proteomes, respectively, and the decay of the former wasquantified through 12 C/( 12 C+ 13 C) ratio maps. We demonstrate time‐dependent imaging of proteomic degradation in mammalian cells under steady‐state conditions and various perturbations, including oxidative stress, cell differentiation, and huntingtin protein aggregation.