title: Proteomics characterization of cytoplasmic and lipid-associated membrane proteins of human pathogen Mycoplasma fermentans M64.
abstract: Mycoplasma fermentans is a potent human pathogen which has been implicated in several diseases. Notably, its lipid-associated membrane proteins (LAMPs) play a role in immunomodulation and development of infection-associated inflammatory diseases. However, the systematic protein identification of pathogenic M. fermentans has not been reported. From our recent sequencing results of M. fermentans M64 isolated from human respiratory tract, its genome is around 1.1 Mb and encodes 1050 predicted protein-coding genes. In the present study, soluble proteome of M. fermentans was resolved and analyzed using two-dimensional gel electrophoresis. In addition, Triton X-114 extraction was carried out to enrich amphiphilic proteins including putative lipoproteins and membrane proteins. Subsequent mass spectrometric analyses of these proteins had identified a total of 181 M. fermentans ORFs. Further bioinformatics analysis of these ORFs encoding proteins with known or so far unknown orthologues among bacteria revealed that a total of 131 proteins are homologous to known proteins, 11 proteins are conserved hypothetical proteins, and the remaining 39 proteins are likely M. fermentans-specific proteins. Moreover, Triton X-114-enriched fraction was shown to activate NF-kB activity of raw264.7 macrophage and a total of 21 lipoproteins with predicted signal peptide were identified therefrom. Together, our work provides the first proteome reference map of M. fermentans as well as several putative virulence-associated proteins as diagnostic markers or vaccine candidates for further functional study of this human pathogen.
authors.affiliation: Institute of Biopharmaceutical Research, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Yi-Chang
authors.initials: YC
authors.lastname: Liu
authors.forename: I-Hsuan
authors.initials: IH
authors.lastname: Lin
authors.forename: Wei-Jen
authors.initials: WJ
authors.lastname: Chung
authors.forename: Wensi S
authors.initials: WS
authors.lastname: Hu
authors.forename: Wailap Victor
authors.initials: WV
authors.lastname: Ng
authors.forename: Chi-Yu
authors.initials: CY
authors.lastname: Lu
authors.forename: Tsung-Yen
authors.initials: TY
authors.lastname: Huang
authors.forename: Hung-Wei
authors.initials: HW
authors.lastname: Shu
authors.forename: Kwang-Jen
authors.initials: KJ
authors.lastname: Hsiao
authors.forename: Shih-Feng
authors.initials: SF
authors.lastname: Tsai
authors.forename: Chuan-Hsiung
authors.initials: CH
authors.lastname: Chang
authors.forename: Chao-Hsiung
authors.initials: CH
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D016680:Genome, Bacterial; D006019:Glycolysis / Q000235:genetics; D006801:Humans; D058635:Lipid-Linked Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D016858:Mycoplasma fermentans / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D016366:Open Reading Frames; D010802:Phylogeny; D017433:Protein Structure, Secondary; D020543:Proteome / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D040901:Proteomics; D017386:Sequence Homology, Amino Acid; D037521:Virulence Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D058635:Lipid-Linked Proteins; D016328:NF-kappa B; D020543:Proteome; D037521:Virulence Factors