title: Mass spectrometry analysis coupled with de novo sequencing reveals amino acid substitutions in nucleocapsid protein from influenza A virus.
abstract: 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.
authors: 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. lizijian@bjmu.edu.cn.
authors: Zijian
authors: Z
authors: Li
authors: Key Laboratory of Zoonosis, Ministry of Education, Jilin University, Changchun 130062, Jilin, China. Wanchunsun@jlu.edu.cn.
authors: Wanchun
authors: W
authors: Sun
authors: Center for Disease Control and Prevention, Changchun 130025, Jilin, China. dl_wu@163.com.
authors: Donglin
authors: D
authors: Wu
authors: Central Laboratory, Jilin University Second Hospital, Changchun 130041, Jilin, China. gaoxiang13@mails.jlu.edu.cn.
authors: Xiang
authors: X
authors: Gao
authors: Central Laboratory, Jilin University Second Hospital, Changchun 130041, Jilin, China. ning668jing@126.com.
authors: Ningning
authors: N
authors: Sun
authors: Central Laboratory, Jilin University Second Hospital, Changchun 130041, Jilin, China. liu_ning@jlu.edu.cn.
authors: Ning
authors: N
authors: Liu
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D019295:Computational Biology; D053118:Influenza A Virus, H1N1 Subtype; D009980:Influenza A virus / Q000145:classification / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000235:genetics*; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D053719:Tandem Mass Spectrometry
chemical_list: D019590:Nucleocapsid Proteins