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Biological Characterization of the Omp1-like Protein from Actinobacillus actinomycetemcomitans

  • Ha, Jung-Hye (Department of Molecular Biology, College of Natural Sciences, Pusan National University) ;
  • Jeong, Mi-Suk (Department of Molecular Biology, College of Natural Sciences, Pusan National University) ;
  • Jo, Wol-Soon (Medical Science Research Center, Dong-A University) ;
  • Jeong, Min-Ho (Medical Science Research Center, Dong-A University) ;
  • Jang, Se-Bok (Department of Molecular Biology, College of Natural Sciences, Pusan National University)
  • Published : 2010.02.20

Abstract

Actinobacillus actinomycetemcomitans is a gram-negative, nonmotile coccobacillus bacterium that is associated with several human diseases, including endocarditis, meningitis, osteomyelitis, subcutaneous abscesses and periodontal diseases. A full-length Omp1-like protein gene from A. actinomycetemcomitans was cloned into a pQE30 vector and overexpressed in Escherichia coli BL21(DE3) cells. The protein revealed sequence homologies to Seventeen kilodalton proteins (Skp) from Pasteurella multocida and E. coli that have been characterized as periplasmic chaperones. This soluble Omp1-like protein was successfully purified to homogeneity for further folding and functional studies. The purity, identity, and conformation of the protein were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis, matrix-assisted laser desorption ionization mass spectrometry, circular dichroism, fluorescence spectroscopic, and differential scanning calorimetric studies. We showed that the protein formed an oligomer larger than a tetramer. We found, further, that it is comprised of mostly $\alpha$-helices and boasts high thermal stability.

Keywords

References

  1. Subbarao, G. V.; van den Berg, B. Journal of Molecular Biology 2006, 360, 750-759. https://doi.org/10.1016/j.jmb.2006.05.045
  2. Ruiz, N.; Maier, E.; Andersen, C.; Benz, R.; Vinas, M. Biophysical Chemistry 2004, 109, 215-227. https://doi.org/10.1016/j.bpc.2003.11.005
  3. Jeong, S. J.; Yee, S. T.; Jo, W. S.; Yu, S. H.; Lee, S. H.; Lim, Y. J.; Yoo, Y. H.; Kim, J. M.; Lee, J. D.; Jeong, M. H. Infection and Immunity 2000, 68, 5132-5138. https://doi.org/10.1128/IAI.68.9.5132-5138.2000
  4. Jo, W. S.; Yee, S. T.; Yoon, S.; Nam, B. H.; Do, E.; Jung, B. S.; Jeong, S. J.; Hong, S. H.; Yoo, Y. H.; Kang, C. D.; Lim, Y. J.; Jeong, M. H; Lee, J. D. Microbiology and Immunology 2006, 50, 535-542. https://doi.org/10.1111/j.1348-0421.2006.tb03823.x
  5. El-Adhami, W.; Kyd, J. M.; Bastin, D. A.; Cripps, A. W. Infection and Immunity 1999, 67, 1935-1942.
  6. Holck, A.; Kleppe, K. Gene 1988, 67, 117-124. https://doi.org/10.1016/0378-1119(88)90014-5
  7. Walton, T. A.; Sousa, M. C. Molecular cell 2004, 15, 367-374. https://doi.org/10.1016/j.molcel.2004.07.023
  8. Gerald, B. CD spectroscopy deconvolution, version 2.1 1997.
  9. Ghaemmaghami, S.; Oas, T. G. Nature structural biology 2001, 8, 879-882. https://doi.org/10.1038/nsb1001-879
  10. Gorziglia, M.; Larrea, C.; Liprandi, F.; Esparza, J. The Journal of general virology 1985, 66, 1889-1900. https://doi.org/10.1099/0022-1317-66-9-1889
  11. Naryzhny, S. N.; Desouza, L. V.; Siu, K. W.; Lee, H. Biochemistry and cell biology 2006, 84, 669-676. https://doi.org/10.1139/O06-037
  12. Tang, P.; Hung, M.; Klostergaard, J. Biochemistry 1996, 35, 8216-8225. https://doi.org/10.1021/bi952182t
  13. Park, S. Bulletin of the Korean Chemical Society 2006, 27, 1885-1887. https://doi.org/10.5012/bkcs.2006.27.11.1885
  14. Hong, J.; Jeong, M. S.; Kim, J. H.; Kim, B. G.; Holbrook, S. R.; Jang, S. B. Bulletin of the Korean Chemical Society 2008, 29, 381-388. https://doi.org/10.5012/bkcs.2008.29.2.381
  15. Qu, J.; Mayer, C.; Behrens, S.; Holst, O.; Kleinschmidt, J. H. Journal of Molecular Biology 2007, 374, 91-105. https://doi.org/10.1016/j.jmb.2007.09.020
  16. Schafer, U.; Beck, K.; Muller, M. The Journal of biological chemistry 1999, 274, 24567-24574. https://doi.org/10.1074/jbc.274.35.24567
  17. Schlapschy, M.; Dommel, M. K.; Hadian, K.; Fogarasi, M.; Korndorfer, I. P.; Skerra, A. Biological chemistry 2004, 385, 137-143. https://doi.org/10.1515/BC.2004.032
  18. Schreiner, H. C.; Sinatra, K.; Kaplan, J. B.; Furgang, D.; Kachlany, S. C.; Planet, P. J.; Perez, B. A.; Figurski, D. H.; Fine, D. H. Proceedings of the National Academy of Sciences of the United States of America 2003, 100, 7295-7300. https://doi.org/10.1073/pnas.1237223100
  19. Arnold, K.; Bordoli, L.; Kopp, J.; Schwede, T. Bioinformatics (Oxford, England) 2006, 22, 195-201. https://doi.org/10.1093/bioinformatics/bti770