Expression and membrane integration of SARS-CoV E protein and its interaction with M protein

Chen, Shih‐Chi; Lo, Shih-Yen; Ma, Hongyang; Li, Huichun

doi:10.1007/s11262-009-0341-6

Cited by 25 publications

(31 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It should be pointed out that HA peptide was fused to the C terminus of the M protein, which is considered to be the essential for M-M interaction and VLP assembly [10,12,18,29,54,55]. Several groups reported that deletion, substitution or extension of the C terminus of the M protein affects interaction with other proteins [6,8,18,[56][57][58]. M protein of most coronaviral species exposes its N-terminus outside the virion, whereas the C-terminal tail is hidden inside the particle (N exo -C endo orientation).…”

Section: Discussionmentioning

confidence: 99%

Novel coronavirus-like particles targeting cells lining the respiratory tract

et al. 2018

View full text Add to dashboard Cite

Virus like particles (VLPs) produced by the expression of viral structural proteins can serve as versatile nanovectors or potential vaccine candidates. In this study we describe for the first time the generation of HCoV-NL63 VLPs using baculovirus system. Major structural proteins of HCoV-NL63 have been expressed in tagged or native form, and their assembly to form VLPs was evaluated. Additionally, a novel procedure for chromatography purification of HCoV-NL63 VLPs was developed. Interestingly, we show that these nanoparticles may deliver cargo and selectively transduce cells expressing the ACE2 protein such as ciliated cells of the respiratory tract. Production of a specific delivery vector is a major challenge for research concerning targeting molecules. The obtained results show that HCoV-NL63 VLPs may be efficiently produced, purified, modified and serve as a delivery platform. This study constitutes an important basis for further development of a promising viral vector displaying narrow tissue tropism.

show abstract

Section: Discussionmentioning

confidence: 99%

Novel coronavirus-like particles targeting cells lining the respiratory tract

et al. 2018

View full text Add to dashboard Cite

show abstract

“…In an alternative minor form, SARS-CoV E protein was shown to be glycosylated on N66, with the C-terminus exposed to the luminal side [80]. A later study using transfected SARS-CoV E protein with an N-terminal Myc-tag confirmed that SARS-CoV E protein was glycosylated co-translationally [83]. Although the two putative TM domains were required for its interaction with the SARS-CoV M protein, the hydrophilic region (60-76) flanking the N66 glycosylation site was dispensable as shown by co-immunoprecipitation experiment [83].…”

Section: Glycosylationmentioning

confidence: 96%

Post-Translational Modifications of Coronavirus Proteins: Roles and Function

2018

View full text Add to dashboard Cite

Post-translational modifications (PTMs) refer to the covalent modifications of polypeptides after they are synthesized, adding temporal and spatial regulation to modulate protein functions. Being obligate intracellular parasites, viruses rely on the protein synthesis machinery of host cells to support replication, and not surprisingly, many viral proteins are subjected to PTMs. Coronavirus (CoV) is a group of enveloped RNA viruses causing diseases in both human and animals. Many CoV proteins are modified by PTMs, including glycosylation and palmitoylation of the spike and envelope protein, N- or O-linked glycosylation of the membrane protein, phosphorylation and ADP-ribosylation of the nucleocapsid protein, and other PTMs on nonstructural and accessory proteins. In this review, we summarize the current knowledge on PTMs of CoV proteins, with an emphasis on their impact on viral replication and pathogenesis. The ability of some CoV proteins to interfere with PTMs of host proteins will also be discussed.

show abstract

“…Incubate the protein samples with magnetic beads by endover-end rotation overnight at 4 °C. 13. Wash the magnetic beads three times with 1 ml modifi ed RIPA buffer.…”

Section: Cells (Frommentioning

confidence: 99%

Studying Coronavirus–Host Protein Interactions

Yang

Hung

et al. 2015

Methods in Molecular Biology

Self Cite

View full text Add to dashboard Cite

To understand the molecular mechanisms of viral replication and pathogenesis, it is necessary to establish the virus-host protein interaction networks. The yeast two-hybrid system is a powerful proteomic approach to study protein-protein interactions. After the identification of specific cellular factors interacting with the target viral protein using the yeast two-hybrid screening system, co-immunoprecipitation and confocal microscopy analyses are often used to verify the virus-host protein interactions in cells. Identification of the cellular factors required for viral survival or eliminating virus infected cells could help scientists develop more effective antiviral drugs. Here we summarize a standard protocol used in our lab to study the coronavirus-host protein interactions, including yeast two-hybrid screening, co-immunoprecipitation, and immunofluorescence microscopy analyses.

show abstract

Expression and membrane integration of SARS-CoV E protein and its interaction with M protein

Cited by 25 publications

References 27 publications

Novel coronavirus-like particles targeting cells lining the respiratory tract

Novel coronavirus-like particles targeting cells lining the respiratory tract

Post-Translational Modifications of Coronavirus Proteins: Roles and Function

Studying Coronavirus–Host Protein Interactions

Contact Info

Product

Resources

About