Detection of receptor-induced glycoprotein conformational changes on enveloped virions by using confocal micro-Raman spectroscopy

Xiaonan Lu; Qian Liu; Javier A Benavides-Montano; Anthony V Nicola; D Eric Aston; Barbara A Rasco; Hector C Aguilar

doi:10.1128/JVI.03220-12

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Detection of receptor-induced glycoprotein conformational changes on enveloped virions by using confocal micro-Raman spectroscopy

Journal article

Open access

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Detection of receptor-induced glycoprotein conformational changes on enveloped virions by using confocal micro-Raman spectroscopy

Xiaonan Lu, Qian Liu, Javier A Benavides-Montano, Anthony V Nicola, D Eric Aston, Barbara A Rasco and Hector C Aguilar

Journal of virology, Vol.87(6), pp.3130-3142

03/2013

DOI: https://doi.org/10.1128/JVI.03220-12

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https://hdl.handle.net/2376/107685

PMCID: PMC3592171

PMID: 23283947

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Abstract

Receptors, Virus - metabolism

Humans

Virion - physiology

Virion - chemistry

Spectrum Analysis, Raman - methods

Virology - methods

Nipah Virus - physiology

Viral Envelope Proteins - chemistry

Viral Envelope Proteins - metabolism

Virus Attachment

Protein Binding

Protein Conformation

Nipah Virus - chemistry

Conformational changes in the glycoproteins of enveloped viruses are critical for membrane fusion, which enables viral entry into cells and the pathological cell-cell fusion (syncytia) associated with some viral infections. However, technological capabilities for identifying viral glycoproteins and their conformational changes on actual enveloped virus surfaces are generally scarce, challenging, and time-consuming. Our model, Nipah virus (NiV), is a syncytium-forming biosafety level 4 pathogen with a high mortality rate (40 to 75%) in humans. Once the NiV attachment glycoprotein (G) (NiV-G) binds the cell receptor ephrinB2 or -B3, G triggers conformational changes in the fusion glycoprotein (F) that result in membrane fusion and viral entry. We demonstrate that confocal micro-Raman spectroscopy can, within minutes, simultaneously identify specific G and F glycoprotein signals and receptor-induced conformational changes in NiV-F on NiV virus-like particles (VLPs). First, we identified reproducible G- and F-specific Raman spectral features on NiV VLPs containing M (assembly matrix protein), G, and/or F or on NiV/vesicular stomatitis virus (VSV) pseudotyped virions via second-derivative transformations and principal component analysis (PCA). Statistical analyses validated our PCA models. Dynamic temperature-induced conformational changes in F and G or receptor-induced target membrane-dependent conformational changes in F were monitored in NiV pseudovirions in situ in real time by confocal micro-Raman spectroscopy. Advantageously, Raman spectroscopy can identify specific protein signals in relatively impure samples. Thus, this proof-of-principle technological development has implications for the rapid identification and biostability characterization of viruses in medical, veterinary, and food samples and for the analysis of virion glycoprotein conformational changes in situ during viral entry.

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Title: Detection of receptor-induced glycoprotein conformational changes on enveloped virions by using confocal micro-Raman spectroscopy
Creators: Xiaonan Lu - Tianjin University of Science and Technology, Tianjin, China
Qian Liu
Javier A Benavides-Montano
Anthony V Nicola
D Eric Aston
Barbara A Rasco
Hector C Aguilar
Publication Details: Journal of virology, Vol.87(6), pp.3130-3142
Academic Unit: Veterinary Microbiology and Pathology, Department of; Paul G. Allen School for Global Animal Health; Food Science, School of
Publisher: United States
Grant note: R21 AI096103 / NIAID NIH HHS AI094329 / NIAID NIH HHS R21 AI094329 / NIAID NIH HHS AI096103 / NIAID NIH HHS
Identifiers: 99900547149901842
Language: English
Resource Type: Journal article