Proteomics

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A photo-oxidation driven proximity labeling strategy enables profiling of mitochondrial proteome dynamics in living cells


ABSTRACT: We herein report a photo-oxidation driven proximity labeling strategy to profile mitochondrial proteome by light dependence in living cells with high spatiotemporal resolution. Taking advantage of organelle-localizable organic photoactivated probe generating reactive species and nucleophilic substrate for proximal proteins oxidation and trapping, mitochondrial proteins were selectively labeled by spatially limited reactions in native environment. Integration of photo-oxidation driven proximity labeling and quantitative proteomics facilitated the plotting of mitochondrial proteome in which up to 310 mitochondrial proteins were identified with the specificity of 64% in HeLa cells. Furthermore, mitochondrial proteome dynamics was deciphered in drug resistant Huh7 and LPS stimulated HMC3 cells which were hard-to-transfect. A number of differential proteins were quantified which were intimately linked to critical processes and provided insights into the related molecular mechanisms of drug resistance and neuroinflammation in the perspective of mitochondria.

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte, Permanent Cell Line Cell, Cell Culture, Macrophage

SUBMITTER: He Wang  

LAB HEAD: He Wang

PROVIDER: PXD036630 | Pride | 2023-03-11

REPOSITORIES: Pride

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A photo-oxidation driven proximity labeling strategy enables profiling of mitochondrial proteome dynamics in living cells.

Wang He H   Wang Zhiting Z   Gao Hang H   Liu Jianhui J   Qiao Zichun Z   Zhao Baofeng B   Liang Zhen Z   Jiang Bo B   Zhang Lihua L   Zhang Yukui Y  

Chemical science 20220928 40


Mapping the proteomic landscape of mitochondria with spatiotemporal precision plays a pivotal role in elucidating the delicate biological functions and complex relationship with other organelles in a variety of dynamic physiological processes which necessitates efficient and controllable chemical tools. We herein report a photo-oxidation driven proximity labeling strategy to profile the mitochondrial proteome by light dependence in living cells with high spatiotemporal resolution. Taking advanta  ...[more]

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