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. 2018 Oct 16;15(1):160.
doi: 10.1186/s12985-018-1073-9.

Mass Spectrometry-Based Investigation of Measles and Mumps Virus Proteome

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Free PMC article

Mass Spectrometry-Based Investigation of Measles and Mumps Virus Proteome

Dora Sviben et al. Virol J. .
Free PMC article

Abstract

Background: Measles (MEV) and mumps virus (MUV) are enveloped, non-segmented, negative single stranded RNA viruses of the family Paramyxoviridae, and are the cause of measles and mumps, respectively, both preventable by vaccination. Aside from proteins coded by the viral genome, viruses are considered to contain host cell proteins (HCPs). The presence of extracellular vesicles (ECVs), which are often co-purified with viruses due to their similarity in size, density and composition, also contributes to HCPs detected in virus preparations, and this has often been neglected. The aim was to identify which virus-coded proteins are present in MEV and MUV virions, and to try to detect which HCPs, if any, are incorporated inside the virions or adsorbed on their outer surface, and which are more likely to be a contamination from co-purified ECVs.

Methods: MUV, MEV and ECVs were purified by ultracentrifugation, hydrophobic interaction chromatography and immunoaffinity chromatography, proteins in the samples were resolved by SDS-PAGE and subjected to identification by MALDI-TOF/TOF-MS. A comparative analysis of HCPs present in all samples was carried out.

Results: By proteomics approach, it was verified that almost all virus-coded proteins are present in MEV and MUV particles. Protein C in MEV which was until now considered to be non-structural viral protein, was found to be present inside the MeV virions. Results on the presence of HCPs in differently purified virus preparations imply that actin, annexins, cyclophilin A, moesin and integrin β1 are part of the virions.

Conclusions: All HCPs detected in the viruses are present in ECVs as well, indicating their possible function in vesicle formation, or that most of them are only present in ECVs. Only five HCPs were constantly present in purified virus preparations, regardless of the purification method used, implying they are likely the integral part of the virions. The approach described here is helpful for further investigation of HCPs in other virus preparations.

Keywords: Extracellular vesicles; Host cell proteins; Mass spectrometry; Measles virus; Mumps virus.

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The authors declare that they have no competing interest.

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Figures

Fig. 1
Fig. 1
SDS-PAGE of MEV sample purified by UC with protein annotations after MALDI-TOF/TOF MS analysis. a, b and c represent three separately prepared samples for which the data are listed in Table 1. CypA – cyclophilin A, HSP – heat shock protein, ni – not identified. Rectangles denote areas where large gel pieces (possibly containing multiple but very faint bands) were excised
Fig. 2
Fig. 2
SDS-PAGE of MUV sample purified by UC with protein annotations after MALDI-TOF/TOF MS analysis. a, b and c represent three separately prepared samples for which the data are listed in Table 1. MVP – major vault protein, TER ATPase – transitional endoplasmatic reticulum ATPase, HSP – heat shock protein, CypA – cyclophilin A, ni – not identified. Rectangles denote areas where large gel pieces (possibly containing multiple but very faint bands) were excised
Fig. 3
Fig. 3
SDS-PAGE of a representative MEV sample purified by HIC with protein annotations after MALDI-TOF/TOF MS analysis. Three separate samples were analysed in total. E1 – eluate with 0.5 M (NH4)2SO4, 50 mM HEPES, E2 – eluate with 50 mM HEPES, MVP – major vault protein, G3BP – galectin-3-binding protein, CypA – cyclophilin A, TER ATPase – transitional endoplasmatic reticulum ATPase, ni – not identified. Rectangles denote areas where large gel pieces (possibly containing multiple but very faint bands) were excised
Fig. 4
Fig. 4
SDS-PAGE of a representative MUV sample purified by HIC with protein annotations after MALDI-TOF/TOF MS analysis. Three separate samples were analysed in total. E1 – eluate with 0.5 M (NH4)2SO4, 50 mM HEPES, E2 – eluate with 50 mM HEPES, MVP – major vault protein, G3BP – galectin-3-binding protein, CypA – cyclophilin A, ni – not identified. Rectangles denote areas where large gel pieces (possibly containing multiple but very faint bands) were excised
Fig. 5
Fig. 5
SDS-PAGE of MUV sample purified by IAC with protein annotations after MALDI-TOF/TOF MS analysis. a and b represent two separately prepared samples for which the data are listed in Table 1. CypA – cyclophilin A, ni – not identified
Fig. 6
Fig. 6
SDS-PAGE of ECVs purified by HIC a, UC b, c, and IAC (d) with protein annotations after MALDI-TOF/TOF MS analysis. MVP – major vault protein, G3BP – galectin-3-binding protein, CypA – cyclophilin A, TER ATPase – transitional endoplasmatic reticulum ATPase, HSP – heat shock protein, GGT – gamma-glutamyl transferase, PDI – protein disulphide-isomerase, BSA – bovine serum albumin, GAPDH – glyceraldehyde-3-phosphate dehydrogenase, GST – glutathione S-transferase, CypA – cyclophilin A, ni – not identified

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