AiM  Vol.3 No.7 , November 2013
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.


Cite this paper
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.
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