Related Experiment Video
Updated: Jun 14, 2026
Separation and Fractionation of Cell Wall and Cell Membrane Proteins from Mycobacterium tuberculosis for Downstream Protein Analysis
Published on: September 26, 2025
Localization of proteins in the cell wall of Mycobacterium avium subsp. paratuberculosis K10 by proteomic analysis
1Department of Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada. jdebuck@ucalgary.ca.
Abstract:
Mycobacterium avium subsp. paratuberculosis is a pathogen which causes a debilitating chronic enteritis in ruminants. Unfortunately, the mechanisms that control M. avium subsp. paratuberculosis persistence during infection are poorly understood and the key steps for developing Johne's disease remain elusive. A proteomic analysis approach, based on one dimensional polyacrylamide gel electrophoresis (SDS-PAGE) followed by LC-MS/MS, was used to identify and characterize the cell wall associated proteins of M. avium subsp. paratuberculosis K10 and an cell surface enzymatic shaving method was used to determine the surface-exposed proteins. 309 different proteins were identified, which included 101 proteins previously annotated as hypothetical or conserved hypothetical. 38 proteins were identified as surface-exposed by trypsin treatment. To categorize and analyze these proteomic data on the proteins identified within cell wall of M. avium subsp. paratuberculosis K10, a rational bioinformatic approach was followed. The analyses of the 309 cell wall proteins provided theoretical molecular mass and pI distributions and determined that 18 proteins are shared with the cell surface-exposed proteome. In short, a comprehensive profile of the M. avium subsp. paratuberculosis K10 cell wall subproteome was created. The resulting proteomic profile might become the foundation for the design of new preventive, diagnostic and therapeutic strategies against mycobacterial diseases in general and M. avium subsp. paratuberculosis in particular.
Insights
Mycobacterium avium subsp. paratuberculosis (MAP) cell wall proteins were identified using proteomics. This study characterized MAP
Area of Science:
- Microbiology
- Proteomics
- Bioinformatics
Background:
- Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease, a chronic enteritis in ruminants.
- Mechanisms of MAP persistence and Johne's disease development are poorly understood.
- Understanding MAP's cell wall and surface proteins is crucial for disease control.
Purpose of the Study:
- To identify and characterize cell wall-associated and surface-exposed proteins of Mycobacterium avium subsp. paratuberculosis K10.
- To create a comprehensive proteomic profile of the MAP K10 cell wall subproteome.
- To provide a foundation for developing new strategies against MAP infections.
Main Methods:
- Proteomic analysis using one-dimensional polyacrylamide gel electrophoresis (SDS-PAGE) and LC-MS/MS.
- Cell surface enzymatic shaving with trypsin treatment to identify surface-exposed proteins.
- Bioinformatic approaches for categorization and analysis of identified proteins.
Main Results:
- 309 different cell wall-associated proteins were identified in MAP K10.
- 101 identified proteins were previously annotated as hypothetical or conserved hypothetical.
- 38 proteins were determined to be surface-exposed, with 18 shared between the cell wall and surface proteomes.
Conclusions:
- A comprehensive proteomic profile of the MAP K10 cell wall subproteome was established.
- This proteomic data provides insights into MAP's cellular structure and potential virulence factors.
- The findings can inform the development of novel preventive, diagnostic, and therapeutic strategies for MAP infections.
