@article{mbs:/content/journal/jgv/10.1099/0022-1317-82-9-2251, author = "Blitvich, Bradley J. and Scanlon, Denis and Shiell, Brian J. and Mackenzie, John S. and Pham, Kim and Hall, Roy A.", title = "Determination of the intramolecular disulfide bond arrangement and biochemical identification of the glycosylation sites of the nonstructural protein NS1 of Murray Valley encephalitis virus", journal= "Journal of General Virology", year = "2001", volume = "82", number = "9", pages = "2251-2256", doi = "https://doi.org/10.1099/0022-1317-82-9-2251", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-82-9-2251", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = "The 12 cysteine residues in the flavivirus NS1 protein are strictly conserved, suggesting that they form disulfide bonds that are critical for folding the protein into a functional structure. In this study, we examined the intramolecular disulfide bond arrangement of NS1 of Murray Valley encephalitis virus and elucidated three of the six cysteine-pairing arrangements. Disulfide linkages were identified by separating tryptic-digested NS1 by reverse-phase high pressure liquid chromatography and analysing the resulting peptide peaks by protein sequencing, amino acid analysis and/or electrospray mass spectrometry. The pairing arrangements between the six amino-terminal cysteines were identified as follows: Cys4–Cys15, Cys55–Cys143 and Cys179–Cys223. Although the pairing arrangements between the six carboxy-terminal cysteines were not determined, we were able to eliminate several cysteine-pairing combinations. Furthermore, we demonstrated that all three putative N-linked glycosylation sites of NS1 are utilized and that the Asn207 glycosylation site contains a mannose-rich glycan.", }