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

Saleh A. Naser, Saisathya Thanigachalam, Natalia Spinelli, Mitra M. Safavi, Najih Naser, Omar Khan

BioSynse, Cary, US.
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, USA.

DOI: 10.4236/aim.2013.37070                

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.

Keywords

Mycobacterium Avium Subspecies Paratuberculosis; Glycolipoprotein; 19 KDa Protein; Post Translational Modification

Conflicts of Interest

The authors declare no conflicts of interest.

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