Dataset Information

Evaluation of a bead-free immunoprecipitation technique coupled with tandem mass spectrometry for identification of plant-virus protein interactions (DeBlasio_JBT_2017)

ABSTRACT: Potato leafroll virus (PLRV) is an aphid-borne, RNA virus in the Luteoviridae that causes significant loss to potato production worldwide. Precision methods to block virus infection in plants or the transmission of a virus by the insect represent new approaches to virus disease management and require an in depth knowledge of the protein interactions used by viruses in their plant hosts and aphid vectors. In this study, we evaluated the feasibility of using an inexpensive, bead-free assay to identify plant proteins that interact with wild-type (WT) and PLRV mutant form. Comparison of host and viral proteins identified as enriched ³2-fold in both WT and mutant experiments using the bead-free method revealed a ~65% overlap with proteins identified as forming high-confident interactions with PLRV using antibody coated-magnetic beads with 19 of these proteins also detected as significant interactions in the bead-free assay. An additional 18 virus-host interactions were only identified in the bead-free assay. Two prey proteins, a 14-3-3 signal transduction protein and malate dehydrogenase 2, were identified as having a weakened or lost association with the mutant form of the virus in both immunoprecipitation assays, showing that the method is sensitive enough to detect quantitative differences between different, yet closely related viral bait proteins. Collectively, our analysis shows that the bead-free platform is a low-cost alternative that can be used by core facilities and other investigators to reproducibly identify plant and viral proteins interacting with virions.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Nicotiana Benthamiana

TISSUE(S): Plant

SUBMITTER: Stacy DeBlasio

LAB HEAD: Michelle Cilia

PROVIDER: PXD006338 | Pride | 2018-10-24

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	A1_NBWT1BSA3PLRV1.mzid	Mzid
	A1_NBWT1BSA3PLRV1.raw	Raw
	A1_NBWT1BSA3PLRV1F395745.mzid_A1_NBWT1BSA3PLRV1_F395745.MGF	Mzid
	A2_NBWT1BSA3PLRV3.mzid	Mzid
	A2_NBWT1BSA3PLRV3.raw	Raw

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Publications

Evaluation of a Bead-Free Coimmunoprecipitation Technique for Identification of Virus-Host Protein Interactions Using High-Resolution Mass Spectrometry.

DeBlasio Stacy L SL Bereman Michael S MS Mahoney Jaclyn J Thannhauser Theodore W TW Gray Stewart M SM MacCoss Michael J MJ Cilia Heck Michelle M

Journal of biomolecular techniques : JBT 20170724 3

Protein interactions between virus and host are essential for viral propagation and movement, as viruses lack most of the proteins required to thrive on their own. Precision methods aimed at disrupting virus-host interactions represent new approaches to disease management but require in-depth knowledge of the identity and binding specificity of host proteins within these interaction networks. Protein coimmunoprecipitation (co-IP) coupled with mass spectrometry (MS) provides a high-throughput way ...[more]

PMID: 28785175

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