Localization of proteins in the cell wall of Mycobacterium avium subsp. paratuberculosis K10 by proteomic analysis

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.

Surface Proteome of “Mycobacterium avium subsp. hominissuis” during the Early Stages of Macrophage Infection

“…Approximately 57% (21/37) of the surface-exposed proteins identified in M. avium subsp. paratuberculosis were also detected in this study (32). Similar results were observed for M. tuberculosis and M. smegmatis, 52% (13/25) and 56% (14/25), respectively (74).…”

“…2, if the scope of reference is expanded to include proteins identified in published studies that comprehensively profile membrane, cell wall, and secreted proteins of M. marinum, M. avium subsp. paratuberculosis, and M. tuberculosis, the results indicate that ϳ90% of the proteins detected in this study are close homologues of proteins previously identified as plasma membrane, secreted, and/or cell wall-associated mycobacterial proteins (30,32,51,54,84,87) (see Table S1 in the supplemental material).…”

“…To assess the relevance of these results, the proteins identified in this screen were cross-referenced against the surface-exposed proteins identified in two published analyses of related mycobacterial species (32,74). Unlike the selective labeling and affinity purification method employed here, the previous work employed either trypsin shaving or cell envelope solubilization to selectively isolate and identify proteins and peptides from the surface of several species of Mycobacterium, including M. avium subsp.…”

Evalution of “indirect fluorescent antibody test‟ as potential screening test for Mycobacterium avium subspecies paratuberculosis using milk of lactating domestic livestock

“…MAP is close to be considered as "zoonotic pathogen" and has high public significance due to large scale presence in the milk of domestic livestock (Singh et al 2014b). Ability of MAP to survive at pasteurization temperature has made milk the primary source of human infection, especially infants, who are increasingly dependent on milk powders (Singh & Vihan, 2004;He & Buck, 2010;Galiero et al, 2015). Crohn"s Disease (CD), a systemic disorder in humans where principal clinico-pathological manifestation is the chronic inflammation of the intestines.…”

Differential Detergent Extraction of Mycobacterium marinum Cell Envelope Proteins Identifies an Extensively Modified Threonine-Rich Outer Membrane Protein with Channel Activity

“…These results indicate that the classical model of outer membrane proteins as (predominantly) being ␤-barrels might not apply to all mycolate outer membrane proteins. Recently, various attempts were made to define proteins in the cell envelope or cell wall fractions of different strains of M. tuberculosis, M. bovis BCG, and Mycobacterium avium (20)(21)(22), but none were dedicated to the specific identification of MOMPs. In this study, we developed a method to specifically enrich for MOM proteins by differential detergent extraction of mycobacterial cell envelopes.…”