Epitope Tagging and Coimmunoprecipitation to Identify Viral Protein Interactors.

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2027. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it

0cells of the map it votes in

0citing papers in PubMed

–field-weighted citation impact

1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

2 authors.

Zhenlan YaoDepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, 90095, USA.

Melody LiDepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, 90095, USA.ORCID 0000-0002-6905-7940

Funding

Functional analysis of host and viral determinants for ZAP inhibitionR01AI158704 · NIAID · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI LI, MELODY · 2021 to 2025

$1.9M

NIAID NIH HHS R01 AI158704

6 · The paper itself

Abstract

Affinity purification-mass spectrometry (AP-MS) is a powerful proteomic approach for dissecting the interaction network between virus and host. Traditional AP-MS employs overexpression of viral proteins as baits to enrich host interactors. However, overexpressed viral proteins may mislocalize to inappropriate cellular compartments and trigger endoplasmic reticulum stress by overwhelming the protein-folding machinery, which leads to false identification of host factors. To overcome these limitations, we introduce an AP-MS strategy based on direct infection with an epitope-tagged chikungunya virus (CHIKV/myc-E2), which we used to successfully uncover two new antiviral factors in CHIKV cellular reservoirs-macrophages. In this protocol, we will describe this technique step by step: (1) design and construction of myc-tagged virus by advanced multi-fragment assembly, (2) in vitro transcription and preparation of infectious myc-tagged virus stocks, and (3) immunoprecipitation of myc-tagged viral protein and its interactome for mass spectrometry analysis. This strategy enables accurate identification of viral interactors in a physiologically relevant context, providing a framework for future proteomic studies using tagged viruses.

Indexed as

Chikungunya virusEpitopesImmunoprecipitationProtein Interaction MappingProteomicsViral ProteinsAnimalsHost-Pathogen InteractionsHumansMass SpectrometryProtein BindingEpitopesViral ProteinsAP-MSChikungunya virusEpitope taggingMacrophageMulti-fragment assemblyTagged virus

Identifiers

PMID42734796

DOI10.3390/v10020089

What OpenQuestion holds

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.