The 19 kDa Protein from Mycobacterium avium subspecies paratuberculosis Is a Glycolipoprotein

Abstract

This study characterizes the 19 kDa protein expressed by Mycobacterium avium subspecies paratuberculosis (MAP) as a glycolipoprotein, providing the foundation for future experiments regarding its antigenicity and role in disease pathogenicity. We have previously shown that a 4.8 kb insert from MAP will produce a 16 kDa recombinant protein when expressed in Escherichia coli and 19 kDa recombinant protein when expressed in M. smegmatis (smeg19K). The difference of 3 kDa in size of these expressed proteins may be related to post translational modifications that occur in Mycobacterium species. We hypothesized that smeg19K is a glycolipoprotein since BLAST analysis revealed approximately 76% amino acid identity between the MAP 19 kDa protein and a known lipoglycoprotein, the 19 kDa protein of M. tuberculosis. This prediction was confirmed by the following positive staining of smeg19K with Sudan Black 4B, a postelectrophoresis dye used to stain for lipids. Smeg19K has also stained positively for glycosylation with the lectin concavalin A, a highly specific stain for mannose residues. As expected, treatment with tunicamycin (an antibiotic known to inhibit N-glycosylation) and treatment with deglycosylation assay (non-specific for mannose), showed no reduction in size of 19 kDa glycolipoprotein.

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S. Naser, S. Thanigachalam, N. Spinelli, M. Safavi, N. Naser and O. Khan, "The 19 kDa Protein from Mycobacterium avium subspecies paratuberculosis Is a Glycolipoprotein," Advances in Microbiology, Vol. 3 No. 7, 2013, pp. 520-528. doi: 10.4236/aim.2013.37070.

Conflicts of Interest

The authors declare no conflicts of interest.

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