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Mass Spectrometry Analysis Coupled with de novo Sequencing Reveals Amino Acid Substitutions in Nucleocapsid Protein from Influenza A Virus

1
Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Ministry of Health/ Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education/ Beijing Key Laboratory of Cardiovascular Receptors Research, Institute of Vascular Medicine, Peking University Third Hospital, Beijing 100191,China
2
Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130062, Jilin, China
3
Center for Disease Control and Prevention, Changchun 130025, Jilin, China
4
Central Laboratory, Jilin University Second Hospital, Changchun 130041, Jilin, China
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2014, 15(2), 2465-2474; https://doi.org/10.3390/ijms15022465
Received: 31 December 2013 / Revised: 26 January 2014 / Accepted: 29 January 2014 / Published: 11 February 2014
(This article belongs to the Special Issue Mass Spectrometry Application in Biology)
Amino acid substitutions in influenza A virus are the main reasons for both antigenic shift and virulence change, which result from non-synonymous mutations in the viral genome. Nucleocapsid protein (NP), one of the major structural proteins of influenza virus, is responsible for regulation of viral RNA synthesis and replication. In this report we used LC-MS/MS to analyze tryptic digestion of nucleocapsid protein of influenza virus (A/Puerto Rico/8/1934 H1N1), which was isolated and purified by SDS poly-acrylamide gel electrophoresis. Thus, LC-MS/MS analyses, coupled with manual de novo sequencing, allowed the determination of three substituted amino acid residues R452K, T423A and N430T in two tryptic peptides. The obtained results provided experimental evidence that amino acid substitutions resulted from non-synonymous gene mutations could be directly characterized by mass spectrometry in proteins of RNA viruses such as influenza A virus. View Full-Text
Keywords: nucleocapsid protein; influenza virus A virus; amino acid substitution; mass spectrometry nucleocapsid protein; influenza virus A virus; amino acid substitution; mass spectrometry
MDPI and ACS Style

Li, Z.; Sun, W.; Wu, D.; Gao, X.; Sun, N.; Liu, N. Mass Spectrometry Analysis Coupled with de novo Sequencing Reveals Amino Acid Substitutions in Nucleocapsid Protein from Influenza A Virus. Int. J. Mol. Sci. 2014, 15, 2465-2474.

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