Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Crystallization and Preliminary X-Ray Crystallographic Analysis of PAS Factor from Vibrio vulnificus

  • Lee, Jun Hyuck (Department of Life Science, Gwangju Institute of Science and Technology) ;
  • Kim, Soo Young (Department of Microbiology, Chonnam National University Medical School) ;
  • Rho, Seong-Hwan (Department of Life Science, Gwangju Institute of Science and Technology) ;
  • Im, Young Jun (Department of Life Science, Gwangju Institute of Science and Technology) ;
  • Kim, Young Ran (Department of Microbiology, Chonnam National University Medical School) ;
  • Kim, Mun-Kyoung (Department of Life Science, Gwangju Institute of Science and Technology) ;
  • Kang, Gil Bu (Department of Life Science, Gwangju Institute of Science and Technology) ;
  • Rhee, Joon Haeng (Department of Microbiology, Chonnam National University Medical School) ;
  • Eom, Soo Hyun (Department of Life Science, Gwangju Institute of Science and Technology)
  • Received : 2005.06.27
  • Accepted : 2005.08.17
  • Published : 2005.12.31
⟨ Previous Next ⟩

Abstract

Plasmid Achromobacter secretion (PAS) factor is a putative secretion factor that induces the secretion of periplasmic proteins. PAS factor from Vibrio vulnificus was crystallized at 294 K by the hanging drop vapor-diffusion method. It was isolated as a monomer during the purification procedures. The native crystal belongs to the F222 space group with unit cell parameters a = 56.1, b = 74.4, $c=80.0{\AA}$, ${\alpha}={\beta}={\gamma}=90^{\circ}$. The crystal was soaked in cryoprotectant containing 1 M NaBr for 1 h for MAD phasing. The diffraction limit of the Br-MAD data set was $1.9{\AA}$ using synchrotron X-ray irradiation at beam line BL-18B at the Photon Factory, Japan.

Keywords

Acknowledgement

Supported by : Ministry of Health and Welfare

References

  1. Finlay, B. B. and Falkow, S. (1997) Common themes in microbial pathogenicity revisited. Microbiol. Mol. Biol. Rev. 61, 136-169
  2. Gray, L. D. and Kreger, A. S. (1989) Detection of Vibrio vulnificus cytolysin in V. vulnificus-infected mice. Toxicon 27, 439-464 https://doi.org/10.1016/0041-0101(89)90206-7
  3. Hampel, A., Labanauskas, M., Connor, P. G., Kirkegard, L., RajBhandary, U. L., et al. (1968) Single crystals of transfer RNA from formylmethionine and phenylalanine transfer RNA's. Science 162, 1384-1387 https://doi.org/10.1126/science.162.3860.1384
  4. Jeong, M. S. and Jang, S. B. (2005) Crystallization and X-ray crystallographic studies of wild-type and mutant tryptophan synthase ${\alpha}$ -subunits from Escherichia coli. Mol. Cells 19, 219-222
  5. Kim, Y. R., Lee, S. E., Kim, C. M., Kim, S. Y., Shin, E. K., et al. (2003) Characterization and pathogenic significance of Vibrio vulnificus antigens preferentially expressed in septicemic patients. Infect. Immun. 71, 5461-5471 https://doi.org/10.1128/IAI.71.10.5461-5471.2003
  6. Kreger, A., DeChatelet, L., and Shirley, P. (1981) Interaction of Vibrio vulnificus with human polymorphonuclear leukocytes: association of virulence with resistance to phagocytosis. J. Infect. Dis. 144, 244-248 https://doi.org/10.1093/infdis/144.3.244
  7. Matthews, B. W. (1968) Solvent content of protein crystals. J. Mol. Biol. 33, 491-497 https://doi.org/10.1016/0022-2836(68)90205-2
  8. McPherson, V. L., Watts, J. A., Simpson, L. M., and Oliver, J. D. (1991) Physiological effects of the lipopolysaccharide of Vibrio vulnificus on mice and rats. Microbios 67, 141-149
  9. Miroux, B. and Walker, J. E. (1996) Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J. Mol. Biol. 260, 289-298 https://doi.org/10.1006/jmbi.1996.0399
  10. Morris, J. G. Jr., Wright, A. C., Simpson, L. M., Wood, P. K., Johnson, D. E., et al. (1987) Virulence of Vibrio vulnificus: association with utilization of transferrin-bound iron, and lack of correlation with levels of cytotoxin or protease production. FEMS Microbiol. Lett. 40, 55-59 https://doi.org/10.1111/j.1574-6968.1987.tb01982.x
  11. Oliver, J. D., Wear, J. E., Thomas, M. B., Warner, M., and Linder, K. (1986) Production of extracellular enzymes and cytotoxicity by Vibrio vulnificus. Diagn. Microbiol. Infect. Dis. 5, 99-111 https://doi.org/10.1016/0732-8893(86)90112-4
  12. Otwinowsky, Z. and Minor, W. (1997) Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307-326 https://doi.org/10.1016/S0076-6879(97)76066-X
  13. Simpson, L. M. and Oliver, J. D. (1987) Ability of Vibrio vulnificus to obtain iron from transferrin and other iron-binding proteins. Curr. Microbiol. 15, 155-157 https://doi.org/10.1007/BF01577265
  14. Terwilliger, T. C. (1999) Reciprocal-space solvent flattening. Acta Cryst. D55, 1863-1871
  15. Terwilliger, T. C. and Berendzen, J. (1999) Automated MAD and MIR structure solution. Acta Cryst. D55, 849-861
  16. Tokugawa, K., Kakitani, M., Ishii, T., Nakamura, K., Masaki, H., et al. (1994a) A novel protein secretion factor from a Vibrio species which operates in Escherichia coli. J. Biotechnol. 35, 69-76 https://doi.org/10.1016/0168-1656(94)90190-2
  17. Tokugawa, K., Ishii, T., Nakamura, K., Masaki, H., and Uozumi, T. (1994b) A model system for the continuous production of a heterologous protein using a novel secretion promoting factor which operates in Escherichia coli. J. Biotechnol. 37, 33-37 https://doi.org/10.1016/0168-1656(94)90199-6
  18. Wright, A. L., Miceli, G. A., Landry, W. L., Christy, J. B., Watkins, W. D., et al. (1993) Rapid identification of Vibrio vulnificus on nonselective media with an alkaline phosphataselabeled oligonucleotide probe. Appl. Environ. Microbiol. 59, 541-546