Identification of 60 secreted proteins for Mycoplasma bovis with secretome assay

• Published in: Microbial pathogenesis Vol. 143; p. 104135
• Main Authors: Zubair, Muhammad, Muhamed, Shafii Abdullahi, Khan, Farhan Anwar, Zhao, Gang, Menghwar, Harish, Faisal, Muhammad, Zhang, Hui, Zhu, Xifang, Rasheed, Muhammad Asif, Chen, Yingyu, Marawan, Marawan A., Chen, Huanchun, Guo, Aizhen
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-06-2020
• Subjects: Antibodies, Bacterial - immunology; Bacterial Proteins - genetics; Bacterial Proteins - immunology; Bacterial Proteins - metabolism; Conserved Sequence - genetics; Electrophoresis, Gel, Two-Dimensional; Genome; Genome, Bacterial - genetics; Genomics; M. bovis; Mass Spectrometry; Mycoplasma bovis - genetics; Mycoplasma bovis - metabolism; Mycoplasma bovis - pathogenicity; Proteomics; Secreted proteins; Secretome; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Virulence; M. bovis; Secretome; Genome; Secreted proteins; Virulence; Proteomics
• ISSN: 0882-4010; 1096-1208
• DOI: 10.1016/j.micpath.2020.104135

Mycoplasma bovis is a risky pathogen mainly responsible for pneumonia and mastitis in cattle. Up to date, its pathogenesis is not clear. Since secreted proteins have a tricky role in M. bovis pathogenesis, this study was designed to systematically reveal M. bovis secretome and potential role in virulence of the pathogen. By using bioinformatics tools, a total of 246 secreted proteins were predicted based on M. bovis genome. Among them, 14 were classical, 154 non-classical and 78 both pathways. Then by using 2-dimensional gel electrophoresis (2-DE) and Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF- MS), 169 proteins were revealed. Of them, 60 were predicted to be secreted including 3 classical, 43 non-classical, and 14 both classical and non-classical. Further 8 proteins (MbovP0038, MbovP0338, MbovP0341, MbovP0520, MbovP0581, MbovP0674, MbovP0693, MbovP0845) were predicted to be virulence-related factors with VFDB. In addition, MbovP0581 (ABC transporter protein) was validated experimentally as secreted in nature and highly immunogenic reacting with sera of cattle experimentally infected with M. bovis. In conclusion, this study might be a crucial step towards a better understanding of pathogenesis and leading to the development of novel diagnostic marker and potent vaccine against M. bovis.
•Secretome of M. bovis was explored.•Combined approach of Genome and Proteome analyses was performed for secreted protein identification.•Bioinformatics tools were applied for prediction analysis.•Identification of 60 secreted proteins.•MbovP0581 was experimentally verified as antigenic secreted protein.