title: Live-cell quantitative imaging of proteome degradation by stimulated Raman scattering.
abstract: 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.
authors.affiliation: Department of Chemistry, Columbia University, New York, NY 10027 (USA).
authors.forename: Yihui
authors.initials: Y
authors.lastname: Shen
authors.forename: Fang
authors.initials: F
authors.lastname: Xu
authors.forename: Lu
authors.initials: L
authors.lastname: Wei
authors.forename: Fanghao
authors.initials: F
authors.lastname: Hu
authors.forename: Wei
authors.initials: W
authors.lastname: Min
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D010649:Phenylalanine / Q000737:chemistry; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D059748:Proteolysis / Q000187:drug effects; D020543:Proteome / Q000032:analysis*; D013059:Spectrum Analysis, Raman
chemical_list: D002247:Carbon Isotopes; D011506:Proteins; D020543:Proteome; D010649:Phenylalanine; D006861:Hydrogen Peroxide
keywords: Raman spectroscopy; SRS microscopy; isotope labeling; protein aggregation; protein degradation
references.citation: Chemphyschem. 2012 Mar;13(4):1054-9
references.pmid: 22368112
references.citation: Science. 2010 Dec 3;330(6009):1368-70
references.pmid: 21127249
references.citation: J Proteome Res. 2011 Dec 2;10(12):5275-84
references.pmid: 22050367
references.citation: Proc Natl Acad Sci U S A. 2013 Jul 9;110(28):11226-31
references.pmid: 23798434
references.citation: Free Radic Biol Med. 2002 Jun 1;32(11):1084-9
references.pmid: 12031893
references.citation: Nat Rev Mol Cell Biol. 2006 Dec;7(12):952-8
references.pmid: 17139335
references.citation: Nat Methods. 2014 Apr;11(4):410-2
references.pmid: 24584195
references.citation: Analyst. 2014 May 21;139(10):2312-7
references.pmid: 24555181
references.citation: Biol Chem. 2008 Mar;389(3):203-9
references.pmid: 18208355
references.citation: Angew Chem Int Ed Engl. 2013 Jul 8;52(28):7266-71
references.pmid: 23703791
references.citation: Anal Chem. 2013 Jan 2;85(1):98-106
references.pmid: 23198914
references.citation: Nature. 2012 Jan 15;481(7382):520-4
references.pmid: 22246323
references.citation: Nature. 2003 Dec 18;426(6968):895-9
references.pmid: 14685250
references.citation: Chem Biol. 2012 Nov 21;19(11):1373-80
references.pmid: 23177192
references.citation: Science. 2011 Feb 11;331(6018):764-8
references.pmid: 21233346
references.citation: Mol Cell Proteomics. 2013 Mar;12(3):638-50
references.pmid: 23242552
references.citation: J Biol Chem. 1958 Mar;231(1):533-45
references.pmid: 13538989
references.citation: Science. 1986 Oct 10;234(4773):179-86
references.pmid: 3018930
references.citation: Nat Rev Mol Cell Biol. 2010 Nov;11(11):777-88
references.pmid: 20944667
references.citation: J Proteome Res. 2009 Jan;8(1):104-12
references.pmid: 18954100
references.citation: Nat Neurosci. 2010 Jul;13(7):897-905
references.pmid: 20543841
references.citation: Nature. 2004 Oct 14;431(7010):805-10
references.pmid: 15483602
references.citation: Annu Rev Neurosci. 2000;23:217-47
references.pmid: 10845064
references.citation: Science. 2008 Dec 19;322(5909):1857-61
references.pmid: 19095943
references.citation: Nature. 2012 Jan 15;481(7382):516-9
references.pmid: 22246326
references.citation: Annu Rev Phys Chem. 2011;62:507-30
references.pmid: 21453061
references.citation: Anal Chem. 2008 Dec 15;80(24):9576-82
references.pmid: 19006335
references.citation: Nat Chem Biol. 2013 Sep;9(9):586-92
references.pmid: 23873212