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TAILS analysis of viral protease 3C substrates

ABSTRACT: Human Hela and murine HL1 cells were cultured and infected in vitro with C3 protease and inactive mutant protease. Cellular proteomes were collected, labeled by isotopic dimethylation and N-terminal peptides were enriched using TALS proteomics.

INSTRUMENT(S): impact II, Q Exactive

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Cell Culture

SUBMITTER: Theo Klein

LAB HEAD: Christopher M. Overall

PROVIDER: PXD008718 | Pride | 2018-03-05

REPOSITORIES: Pride

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Publications

N-Terminomics TAILS Identifies Host Cell Substrates of Poliovirus and Coxsackievirus B3 3C Proteinases That Modulate Virus Infection.

Jagdeo Julienne M JM Dufour Antoine A Klein Theo T Solis Nestor N Kleifeld Oded O Kizhakkedathu Jayachandran J Luo Honglin H Overall Christopher M CM Jan Eric E

Journal of virology 20180328 8

Enteroviruses encode proteinases that are essential for processing of the translated viral polyprotein. In addition, viral proteinases also target host proteins to manipulate cellular processes and evade innate antiviral responses to promote replication and infection. Although some host protein substrates of enterovirus proteinases have been identified, the full repertoire of targets remains unknown. We used a novel quantitative <i>in vitro</i> proteomics-based approach, termed <u>t</u>erminal < ...[more]

PMID: 29437971

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Project description:The N-terminome in EBV-B cells from a human patient homozygous for an W580S mutation in MALT1 were analyzed by TMT10-plex TAILS and compared to EBV-B cells from heterozygous subjects under resting and PMA/ionomycin stimulation for 2 and 4 h.

2018-10-03 | PXD006723 | Pride

MitoTAILS: N-terminomics profiling of mitochondrial proteolysis

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mito-TAILS analysis of EPEC infection

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Identification of novel natural substrates of fibroblast activation protein by differential degradomics and proteomics (TAILS dataset)

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2018-12-19 | PXD004757 | Pride

Insight into the Human Pathodegradome of the V8 protease from Staphylococcus aureus

Project description:The major human pathogen Staphylococcus aureus possesses ten extracellular proteases that are central to the virulence process. While some host targets of these and other bacterial proteases are known, a major bottleneck to the study of their function is the low-throughout nature of substrate identification. Rapid identification of proteolytic cleavage events in complex substrates has, however, been rendered possible by recent developments in the proteomics sub discipline of N-terminomics. Thus, herein we perform the first study of bacterial protease-host interactions, combining the S. aureus V8 protease and the infection relevant substrate human serum. In so doing, we identified ~90 host-protein targets of V8, including previously known and novel targets. Amongst these were complement components (C1r, C1s, C3,C4b, C4BPA, C5, C6, C8G, C9, CfB, CfH, and ficolin), iron sequestration and transport proteins (ceruloplasmin, serotransferrin, hemopexin, haptoglobin), clotting cascade proteins (plasminogen, kallikrein proteins, prothrombin, factors X and XII), and multiple host protease inhibitors (α2M and members of the ITIH and SERPIN families). N-terminomics also provided insight into the disruption of host protein function by highlighting specific target regions of host proteins, where V8 cleaved within peptidase and sushi domains of complement proteins, and, in the case of host protease inhibitors, cleavage occurred predominantly outside their protease-trapping motifs. Collectively, our data highlight the potential for further application of N-terminomics for the discovery of bacterial protease substrates in other host niches, and provides unique insight into the role of the V8 protease in S. aureus pathogenesis.

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A human endogenous retrovirus encoded protease potentially cleaves numerous cellular proteins

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