Transmissible gastroenteritis virus: Identification of M protein-binding peptide ligands with antiviral and diagnostic potential

Zhang, Hao; Zarlenga, Dante S.; Sestak, Karol; Suo, Siqingaowa; Ren, Xiaofeng

doi:10.1016/j.antiviral.2013.06.015

Cited by 25 publications

(18 citation statements)

References 42 publications

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“…Given the novelty of the virus, they were unable to compare it to other assays currently in use. Zou et al [35] use techniques similar to those developed here, i.e., peptide display, to target the M protein of TGEV in developing an ELISA-based diagnostic test. In this case, the sensitivity of the ELISA exceeded that When the phage-mediated ELISA and antibody ELISA were compared to RT-PCR which targeted a 208-base pair fragment of the S gene, the RT-PCR was most sensitive of all assays tested.…”

Section: Resultsmentioning

confidence: 96%

Phage display for identifying peptides that bind the spike protein of transmissible gastroenteritis virus and possess diagnostic potential

et al. 2015

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The spike (S) protein of porcine transmissible gastroenteritis virus (TGEV) is located within the viral envelope and is the only structural protein that possesses epitopes capable of inducing virus-neutralizing antibodies. Among the four N-terminal antigenic sites A, B, C, and D, site A and to a lesser extent site D (S-AD) induce key neutralizing antibodies. Recently, we expressed S-AD (rS-AD) in recombinant form. In the current study, we used the rS-AD as an immobilized target to identify peptides from a phage-display library with application for diagnosis. Among the 9 phages selected that specifically bound to rS-AD, the phage bearing the peptide TLNMHLFPFHTG bound with the highest affinity and was subsequently used to develop a phage-based ELISA for TGEV. When compared with conventional antibody-based ELISA, phage-mediated ELISA was more sensitive; however, it did not perform better than semi-quantitative RT-PCR, though phage-mediated ELISA was quicker and easier to set up.

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Section: Resultsmentioning

confidence: 96%

Phage display for identifying peptides that bind the spike protein of transmissible gastroenteritis virus and possess diagnostic potential

et al. 2015

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“…The ascites were centrifuged at 12,000 rpm for 10 min, and the supernatant was purified by the caprylic acid-ammonium sulfate method as previously described [13]. Biopanning with a 12-mer phage display peptide library was done according to the manufacturer's instructions with minor modifications [14]. During first to third rounds of panning, the purified MAb-5E12 was incubated with the crude phage library or with eluted phage (1.5 9 10 11 pfu).…”

Section: Biopanningmentioning

confidence: 99%

Putative phage-display epitopes of the porcine epidemic diarrhea virus S1 protein and their anti-viral activity

Cao

Gao

et al. 2015

Virus Genes

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Porcine epidemic diarrhea virus (PEDV) is a pathogen of swine that causes severe diarrhea and dehydration resulting in substantial morbidity and mortality in newborn piglets. Phage display is a technique with wide application, in particular, the identification of key antigen epitopes for the development of therapeutic and diagnostic reagents and vaccines. To identify antigen epitopes with specificity for PEDV, a monoclonal antibody (MAb-5E12) against the immunodominant region of the PEDV Spike protein (S1) was used as the target for biopanning a 12-mer phage display, random peptide library. After multiple rounds of biopanning and stringent washing, three phage-displayed peptides, designated L, W and H, were identified that recognize MAb-5E12. Sequence analysis showed that the one or more of the peptides exhibited partial sequence similarity to the native S1 sequence 'MQYVYTPTYYML' (designated peptide M) at position 201-212. In combination with software analysis for the prediction of B cell epitopes, aa 201-212 exhibited characteristics of a linear epitope on the PEDV S1 protein. In contrast to peptide M, a consensus motif 'PxxY' was identified on both peptides L and W, and on the S1 protein, but not on peptide H. Peptide M and the MAb-5E12-recognizing peptides L and W significantly inhibited the adsorption of PEDV on the cell surface as monitored through plaque-reduction assays. Furthermore, data from real-time PCR and indirect immunofluorescence assays were consistent with the ability of peptides M, L and W to block viral protein expression and thereby function as antiviral agents for PEDV.

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“…As an important structural protein of TGEV particles, the M protein is located in the lipid envelop and associates with the Golgi complex in the cell, suggesting that the M protein plays a pivotal role in virus assembly and budding. Additional evidence indicates that the M protein is an indispensable part for the replication of virus particles in host cells, which may also be used for virus-specific diagnostics and treatment (Cologna and Hogue, 1998;De Haan et al, 1998;Zou et al, 2013). However, there have been few reports regarding the interaction between the TGEV M protein and intestinal cells.…”

Section: Introductionmentioning

confidence: 99%

EIF4A2 interacts with the membrane protein of transmissible gastroenteritis coronavirus and plays a role in virus replication

Song

Yang

Wang

et al. 2019

Research in Veterinary Science

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Transmissible gastroenteritis coronavirus (TGEV) is enteropathogenic coronavirus that causes diarrhea in pigs, and is associated with high morbidity and mortality in sucking piglets. The TGEV membrane (M) protein is a decisive protein for the proliferation of viral proteins, and is associated with virus assembly and budding. To identify the cellular proteins that interact with the TGEV M protein, yeast two-hybrid screening was employed, and seven cellular proteins were identified M-binding partners. Using the GST pull-down approach and a CO-IP assay, the M protein was found to interact with porcine intestinal cells via eukaryotic translation initiation factor 4-alpha (EIF4A2), an essential component of the cellular translational machinery. Additionally, confocal microscopy revealed that EIF4A2 and M were colocalized in the cytoplasm. Furthermore, the function of EIF4A2 in intestinal cells during TGEV infection was examined. A knockdown of EIF4A2 by siRNA markedly decreased M protein proliferation and TGEV replication in target cells. Thus demonstrating that EIF4A2 plays a significant role in TGEV replication. The present study provides mechanistic insight into the interaction between the TGEV M protein and intestinal cells which contributes to the understanding of coronavirus replication and may be useful for the development of novel therapeutic strategies for TGEV infection.

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Transmissible gastroenteritis virus: Identification of M protein-binding peptide ligands with antiviral and diagnostic potential

Cited by 25 publications

References 42 publications

Phage display for identifying peptides that bind the spike protein of transmissible gastroenteritis virus and possess diagnostic potential

Phage display for identifying peptides that bind the spike protein of transmissible gastroenteritis virus and possess diagnostic potential

Putative phage-display epitopes of the porcine epidemic diarrhea virus S1 protein and their anti-viral activity

EIF4A2 interacts with the membrane protein of transmissible gastroenteritis coronavirus and plays a role in virus replication

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