A comprehensive proteomic analysis of Mycobacterium bovis bacillus Calmette–Guérin using high resolution Fourier transform mass spectrometry

• Published in: Journal of proteomics Vol. 77; pp. 357 - 371
• Main Authors: Zheng, Jianhua, Liu, Liguo, Wei, Candong, Leng, Wenchuan, Yang, Jian, Li, Weijun, Wang, Jin, Jin, Qi
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 21.12.2012; Elsevier
• Subjects: Bacterial Proteins - immunology; Bacterial Proteins - metabolism; BCG Vaccine - immunology; BCG Vaccine - metabolism; BCG Vaccine - therapeutic use; Biological and medical sciences; Comprehensive proteomic analysis; Diverse techniques; Fundamental and applied biological sciences. Psychology; LTQ Orbitrap Velos mass spectrometer; Mass Spectrometry - methods; Molecular and cellular biology; Mycobacterium bovis - immunology; Mycobacterium bovis - metabolism; Mycobacterium bovis bacillus Calmette–Guérin; Proteome - immunology; Proteome - metabolism; Proteomics - methods; Pseudogene; RT-PCR; Tuberculosis - immunology; Tuberculosis - metabolism; Tuberculosis - prevention & control; Vaccination; LTQ Orbitrap Velos mass spectrometer; Pseudogene; Comprehensive proteomic analysis; Mycobacterium bovis bacillus Calmette–Guérin; RT-PCR; High resolution; Mycobacterium bovis; Bacillaceae; Bacillales; Mycobacteriales; Analysis; Bacillus; Proteomics; Mycobacteriaceae; Calmette―Guérin; Mycobacterium bovis bacillus; Bacteria; Actinomycetes; Reverse transcription polymerase chain reaction; Mass spectrometry
• ISSN: 1874-3919; 1876-7737
• DOI: 10.1016/j.jprot.2012.09.010

Since 1921, Mycobacterium bovis bacillus Calmette–Guérin (BCG) has been recognized as an important vaccine to prevent tuberculosis worldwide. Nonetheless, a global analysis of BCG proteome has not been clearly investigated. In this study, we performed an in-depth proteomic analysis of BCG under an in vitro cultivation condition using SDS-PAGE and high resolution Fourier transform mass spectrometry. In total, 3434 proteins (35,259 unique peptides) including 512 transmembrane proteins were identified, covering ~87% of the predicted BCG proteome. Seven pseudogene protein products were also obtained and validated by RT-PCR at gene transcript level. Additionally, translational start sites of 832 proteins were confirmed and 186 were extended using N-terminus-derived peptides. The physicochemical characteristics of all identified proteins were determined. Some predominant proteins, including PE and PPE family proteins, lipoproteins, heat shock proteins, transport proteins and low molecular weight protein antigens, are discussed, which represent potential prominent antigens in the humoral and cellular immune response. This study represents the most comprehensive BCG proteome to date, which will likely facilitate the design of vaccination and immunodiagnostic strategies against TB. [Display omitted] ► We performed an in-depth proteomic analysis of BCG using 1-DE and accurate MS. ► In total, 3434 proteins, including 512 membrane proteins, were identified. ► The results represent ~87% of the predicted BCG proteome. ► Seven pseudogenes and a number of extending genes were also detected. ► Some proteins represent potential candidates for vaccination and immunodiagnosis.