Activity-based proteome profiling of hepatoma cells during Hepatitis C virus replication using protease substrate probes

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DOIResolve DOI: http://doi.org/10.1021/pr900788a
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TypeArticle
Journal titleJournal of Proteome Research
Volume9
Issue2
Pages912923; # of pages: 12
Subjectactivity-based protein profiling; amino acid coupled quinolimine methide probes; carboxylesterase 1; hepatitis C; hydrolases; liver; proteases; protein disulfide isomerase
AbstractActivity-based protein profiling (ABPP) offers direct insight into changes in catalytic activity of enzyme classes in complex proteomes, rather than protein or transcript abundance. Here, ABPP was performed in Huh7 hepatoma cell lines with a group of ABPP probes composed of an N-acetylated amino acid, that mimic the P1 position in protease peptide substrates. Five different probes bearing distinct amino acids (Ser, Thr, Phe, Glu and His) labeled 54 differentially active proteins, including proteases, other hydrolases, oxidoreductases and isomerases. Four of the six protease families were targeted based on their P1 substrate preferences. The broader specificity of the labeling observed could be explained by the substrate-based targeting nature and the electrophilic properties of the ABPP probes. When applied to Huh7 cells stably replicating hepatitis C virus (HCV) subgenomic replicon RNA, four proteins showed reduced activity, while three proteins had increased activity during HCV replication. These differentially active hits included carboxylesterase 1, cathepsin D, HSP105, protein disulfide isomerise 1 and A6, chaperonin containing TCP1 and isochorismatase domain containing 1, which demonstrated substrate preferences by being labeled by specific substrate probes. This illustrates the broader activitybased profiling capabilities of these substrate-based probes to reveal novel enzyme candidates and their potential roles during HCV replication.
Publication date
LanguageEnglish
AffiliationNRC Steacie Institute for Molecular Sciences; National Research Council Canada
Peer reviewedYes
NPARC number17104546
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Record identifiere090e08c-f4e8-4585-a807-f25105fad5bc
Record created2011-03-15
Record modified2016-05-09
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