Identification of mumps virus protein and lipid composition by mass spectrometry
- PDF / 3,162,336 Bytes
- 10 Pages / 595.276 x 790.866 pts Page_size
- 112 Downloads / 203 Views
RESEARCH
Open Access
Identification of mumps virus protein and lipid composition by mass spectrometry Marija Brgles1,2*, Maximilian Bonta3, Maja Šantak1,2, Maja Jagušić1,2, Dubravko Forčić1,2, Beata Halassy1,2, Günter Allmaier3 and Martina Marchetti-Deschmann3
Abstract Background: Mumps virus is a negative-sense, single stranded RNA virus consisting of a ribonucleocapsid core enveloped by a lipid membrane derived from host cell, which causes mumps disease preventable by vaccination. Since virus lipid envelope and glycosylation pattern are not encoded by the virus but dependent on the host cell at least to some extent, the aim of this work was to analyse L-Zagreb (L-Zg) mumps virus lipids and proteins derived from two cell types; Vero and chicken embryo fibroblasts (CEF). Jeryl Lynn 5 (JL5) mumps strain lipids were also analysed. Methods: Virus lipids were isolated by organic phase extraction and subjected to 2D-high performance thin layer chromatography followed by lipid extraction and identification by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Virus samples were also subjected to gel electrophoresis under denaturating conditions and protein bands were excised, in-gel trypsinized and identified by MS as well as tandem MS. Results: Results showed that lipids of both mumps virus strains derived from Vero cells contained complex glycolipids with up to five monosaccharide units whereas the lipid pattern of mumps virus derived from CEF was less complex. Mumps virus was found to contain expected structural proteins with exception of fusion (F) protein which was not detected but on the other hand, V protein was detected. Most interesting finding related to the mumps proteins is the detection of several forms of nucleoprotein (NP), some of which appear to be C-terminally truncated. Conclusions: Differences found in lipid and protein content of mumps virus demonstrated the importance of detailed biochemical characterization of mumps virus and the methodology described here could provide a means for a more comprehensive quality control in vaccine production. Keywords: Mumps virus, Proteome, Lipidome, Vero cells, Chicken embryo fibroblasts, Mass spectrometry
Background Mumps virus is a nonsegmented, negative stranded RNA virus of the family Paramyxoviridae, subfamily Paramyxovirinae, genus Rubulavirus that causes mumps disease. Mumps virions are pleomorphic particles ranging from 100 to 800 nm in size [1], consisting of a helical ribonucleocapsid core surrounded by a host cell-derived lipid envelope. Full-length genomic RNA consisting of 15384 nucleotides contains 7 tandemly linked transcription * Correspondence: [email protected] 1 University of Zagreb, Centre for Research and Knowledge Transfer in Biotechnology, Rockefellerova 10, HR-10000 Zagreb, Croatia 2 Center of Excellence for Viral Immunology and Vaccines, CERVirVac, Rijeka, Zagreb, Croatia Full list of author information is available at the end of the article
units that encode open reading frames for the nucleoprotein (NP), phosph
Data Loading...
