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000864979 1001_ $$0P:(DE-HGF)0$$aWeng, Samuel SH$$b0
000864979 245__ $$aSensitive determination of proteolytic proteoforms in limited microscale proteome samples
000864979 260__ $$aBethesda, Md.$$bThe American Society for Biochemistry and Molecular Biology$$c2019
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000864979 520__ $$aProtein N termini unambiguously identify truncated, alternatively translated or modified proteoforms with distinct functions and reveal perturbations in disease. Selective enrichment of N-terminal peptides is necessary to achieve proteome-wide coverage for unbiased identification of site-specific regulatory proteolytic processing and protease substrates. However, many proteolytic processes are strictly confined in time and space and therefore can only be analyzed in minute samples that provide insufficient starting material for current enrichment protocols. Here we present High-efficiency Undecanal-based N Termini EnRichment (HUNTER), a robust, sensitive and scalable method for the analysis of previously inaccessible microscale samples. HUNTER achieved identification of >1000 N termini from as little as 2 μg raw HeLa cell lysate. Broad applicability is demonstrated by the first N-terminome analysis of sorted human primary immune cells and enriched mitochondrial fractions from pediatric cancer patients, as well as protease substrate identification from individual Arabidopsis thaliana wild type and Vacuolar Processing Enzyme-deficient mutant seedlings. We further implemented the workflow on a liquid handling system and demonstrate the feasibility of clinical degradomics by automated processing of liquid biopsies from pediatric cancer patients.
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000864979 536__ $$0G:(EU-Grant)639905$$aProPlantStress - Proteolytic processing in plant stress signal transduction and responses to abiotic stress and pathogen attack (639905)$$c639905$$fERC-2014-STG$$x1
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000864979 7001_ $$0P:(DE-Juel1)167325$$aDemir, Fatih$$b1
000864979 7001_ $$0P:(DE-HGF)0$$aErgin, Enes K.$$b2
000864979 7001_ $$0P:(DE-Juel1)164348$$aDirnberger, Sabrina$$b3
000864979 7001_ $$0P:(DE-HGF)0$$aUzozie, Anuli$$b4
000864979 7001_ $$0P:(DE-Juel1)159510$$aTuscher, Domenic$$b5
000864979 7001_ $$0P:(DE-HGF)0$$aNierves, Lorenz$$b6
000864979 7001_ $$0P:(DE-HGF)0$$aTsui, Janice$$b7
000864979 7001_ $$0P:(DE-Juel1)162356$$aHuesgen, Pitter F.$$b8$$eCorresponding author
000864979 7001_ $$00000-0003-1171-5864$$aLange, Philipp F.$$b9$$eCorresponding author
000864979 773__ $$0PERI:(DE-600)2071375-7$$a10.1074/mcp.TIR119.001560$$gp. mcp.TIR119.001560 -$$n11$$p2335-2347$$tMolecular & cellular proteomics$$v18$$x1535-9484$$y2019
000864979 8564_ $$uhttps://juser.fz-juelich.de/record/864979/files/2335.full-2.pdf$$yPublished on 2019-08-30. Available in OpenAccess from 2020-08-30.
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