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Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis
Published on: May 23, 2021
Outer membrane proteome analysis of Indian strain of Pasteurella multocida serotype B:2 by MALDI-TOF/MS analysis
A Prasannavadhana1, Santosh Kumar1, Prasad Thomas2
1Division of Veterinary Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh 243122, India.
Abstract:
Identification of outer membrane proteins (OMPs) is important to understand the bacteria structure and function, host-pathogen interaction, development of novel vaccine candidates, and diagnostic antigens. But till now the key antigens of P. multocida B:2 isolate causing haemorrhagic septicaemia (HS) in animals are not clearly defined. In this study, P52 strain of P. multocida serotype B:2 was grown in vitro under iron-rich and iron-limited condition. The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis. In total, 22 proteins were identified, of which 7 were observed exclusively under iron-limited condition. Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition. Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2. Further functional in vivo study of the proteins identified are required to explore the utility of these proteins in developing diagnostics and vaccine against HS.
Insights
This study identified key outer membrane proteins (OMPs) from *P. multocida* B:2, crucial for understanding haemorrhagic septicaemia (HS) and developing new vaccines and diagnostics.
Area of Science:
- Microbiology
- Bacterial Pathogenesis
- Immunology
Background:
- Outer membrane proteins (OMPs) are vital for bacterial structure, function, and host interactions.
- Defining key antigens of *P. multocida* B:2 is essential for developing vaccines and diagnostics against haemorrhagic septicaemia (HS).
Purpose of the Study:
- To identify outer membrane proteins (OMPs) of *P. multocida* B:2, particularly under iron-limited conditions.
- To discover potential novel antigens for diagnostics and vaccines against HS.
Main Methods:
- Cultured *P. multocida* P52 strain under iron-rich and iron-limited conditions.
- Extracted OMPs using sarkosyl method, followed by SDS-PAGE.
- Identified proteins using MALDI-TOF/MS analysis.
Main Results:
- Identified a total of 22 OMPs, with 7 exclusively found under iron-limited conditions.
- Highlighted several high molecular weight proteins involved in iron acquisition (e.g., TbpA, HgbA).
- Reported novel hypothetical proteins potentially unique to *P. multocida* B:2.
Conclusions:
- The identified OMPs, especially those related to iron acquisition and novel hypothetical proteins, are potential candidates for HS diagnostics and vaccines.
- Further in vivo functional studies are necessary to validate their utility.
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