Bioinformatics and Molecular Biological Characterization of a Hypothetical Protein SAV1226 as a Potential Drug Target for Methicillin/Vancomycin- Staphylococcus aureus Infections

Authors: Nichole Haag, Kimberly Velk, Tyler McCune, Chun Wu

Abstract:

Methicillin/multiple-resistant Staphylococcus aureus (MRSA) are infectious bacteria that are resistant to common antibiotics. A previous in silico study in our group has identified a hypothetical protein SAV1226 as one of the potential drug targets. In this study, we reported the bioinformatics characterization, as well as cloning, expression, purification and kinetic assays of hypothetical protein SAV1226 from methicillin/vancomycin-resistant Staphylococcus aureus Mu50 strain. MALDI-TOF/MS analysis revealed a low degree of structural similarity with known proteins. Kinetic assays demonstrated that hypothetical protein SAV1226 is neither a domain of an ATP dependent dihydroxyacetone kinase nor of a phosphotransferase system (PTS) dihydroxyacetone kinase, suggesting that the function of hypothetical protein SAV1226 might be misannotated on public databases such as UniProt and InterProScan 5.

Keywords: Dihydroxyacetone kinase, essential genes, Methicillin-resistant Staphylococcus aureus, drug target.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1107173

References:

[1] Voyich, J. et al. (2006) J. Infect. Dis., 194(12):1761-1770.
[2] CDC (2007) MRSA: Methicillin-resistant Staphylococcus aureus in Healthcare Settings.
[3] Klevens, R. et al. (2007), J. Am. Med. Assn., 298(15):1763-1771.
[4] Mulligan, E. et al. (1993). Am. J. Med. 94: 313-328.
[5] Cunha, A. (2005), Clin. Microbiol. Infec., 11, (s4): 33–42,
[6] Walsh, T. et al. (2009). Sci. Am. 301(1): 44-51.
[7] Weinstein, R. A. (2001) Emerg Infect Dis. 7(2):188–192.
[8] Nathan, C. (2004) Nature, 431: 899–902.
[9] Tenover, F. C. (2006) Am. J. Med.119 (6 Suppl 1):S3–S10.
[10] Barrett C. et. al. (2003) Curr. Opin. Biotech. 14: 621–626.
[11] Walsh, T. (2003) Nature Rev. Microbiol. 1:65–70.
[12] Haag, N. et al. (2012), International Journal on Advances in Life Sciences, 4 (1&2):21-32.
[13] Unpublished result.
[14] Daniel, R. et al. (1995) J. Bacteriol. 177:4392–4401
[15] Gutknecht, R. et al. (2001) EMBO J., 20(10):2480-6.
[16] http://www.phantome.org/PhageSeed/SubsysEditor.cgi?page=ShowSubs ystem&subsystem=Dihydroxyacetone_kinases (11/26/2014)
[17] http://www.ncbi.nlm.nih.gov/protein/15924216 (11/26/2014)
[18] http://www.genome.jp/dbget-bin/www_bget?sav:SAV1226(11/26/2014)
[19] http://www.uniprot.org/uniprot/Q99UP2 (11/26/2014)
[20] http://www.ebi.ac.uk/Tools/pfa/iprscan (11/26/2014)
[21] Altschul, F. et al. (1997), Nucleic Acids Res. 25:3389-3402.8