title: Experimental Validation of Bacillus anthracis A16R Proteogenomics.
abstract: Anthrax, caused by the pathogenic bacterium Bacillus anthracis, is a zoonosis that causes serious disease and is of significant concern as a biological warfare agent. Validating annotated genes and reannotating misannotated genes are important to understand its biology and mechanisms of pathogenicity. Proteomics studies are, to date, the best method for verifying and improving current annotations. To this end, the proteome of B. anthracis A16R was analyzed via one-dimensional gel electrophoresis followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). In total, we identified 3,712 proteins, including many regulatory and key functional proteins at relatively low abundance, representing the most complete proteome of B. anthracis to date. Interestingly, eight sequencing errors were detected by proteogenomic analysis and corrected by resequencing. More importantly, three unannotated peptide fragments were identified in this study and validated by synthetic peptide mass spectrum mapping and green fluorescent protein fusion experiments. These data not only give a more comprehensive understanding of B. anthracis A16R but also demonstrate the power of proteomics to improve genome annotations and determine true translational elements.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Zhiqi
authors.initials: Z
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Engineering Research Center for Protein Drugs, National Center for Protein Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Haidian District, Beijing 100850, China.|Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuchang District, Wuhan 430071, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Institute of Computing Technology, Chinese Academy of Sciences, 6 Kexueyuan Nanlu, Haidian District, Beijing 100190, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Engineering Research Center for Protein Drugs, National Center for Protein Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Haidian District, Beijing 100850, China.
authors.forename: Yanchang
authors.initials: Y
authors.lastname: Li
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Engineering Research Center for Protein Drugs, National Center for Protein Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Haidian District, Beijing 100850, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Dongshu
authors.initials: D
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Xiankai
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Erling
authors.initials: E
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Engineering Research Center for Protein Drugs, National Center for Protein Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Haidian District, Beijing 100850, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Chang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Xu
authors.affiliation: Institute of Computing Technology, Chinese Academy of Sciences, 6 Kexueyuan Nanlu, Haidian District, Beijing 100190, China.
authors.forename: Simin
authors.initials: S
authors.lastname: He
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Engineering Research Center for Protein Drugs, National Center for Protein Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Haidian District, Beijing 100850, China.|Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences, 185 Donghu Road, Wuchang District, Wuhan 430071, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, China.
authors.forename: Hengliang
authors.initials: H
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D001408:Bacillus anthracis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D008969:Molecular Sequence Data; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D013171:Spores, Bacterial
chemical_list: D001426:Bacterial Proteins; D020543:Proteome