Mass spectrometric identification and characterisation of the nucleocapsid protein of Menangle virus.

The recent emergence of novel viruses requires reliable methodology for their identification and confirmation both on a cellular and molecular level. Mass spectrometry offers a suitable approach for the identification and characterisation of viral proteins and its application is demonstrated in this study. Menangle virus is a previously unclassified member of the family Paramyxoviridae isolated in Australia in 1997. Menangle virus caused disease in pregnant pigs, and like the other newly emergent Hendra, Nipah and Tioman viruses, appears to be a virus of fruit bats (flying foxes) in the genus Pteropus. The 61 kDa gel-purified protein isolated from cell-associated Menangle virus ribonucleoprotein (RNP) was identified as the nucleocapsid protein (NP) by peptide mapping, mass spectrometry and amino acid sequencing. Over 69% of the amino acid sequence was obtained and found to be identical with that derived from gene analysis (Virology, 283 (2001), 358). The first residue of the mature NP was found to be serine (second residue in the gene derived amino acid sequence). The NP was found to be acetylated at the N-terminus (at Ser-2) and appears to be not modified by phosphorylation.