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http://dx.doi.org/10.4014/kjmb.1205.05013

Cloning, Expression, and Polymerization Assay of FtsZ Protein from Staphylococcus aureus  

Son, Sang Hyeon (Department of Bio & Nano Chemistry, Kookmin University)
Lee, Dong Yun (Seoul Science High School)
Kim, Ye Jun (Seoul Science High School)
Ko, Sooho (Seoul Science High School)
Cho, Seong Jun (Seoul Science High School)
Jung, Hyo Cheol (Seoul Science High School)
Lee, Hyung Ho (Department of Bio & Nano Chemistry, Kookmin University)
Publication Information
Microbiology and Biotechnology Letters / v.40, no.3, 2012 , pp. 274-277 More about this Journal
Abstract
Cytokinesis is the final stage of cell division, dividing one mother cell into two daughter cells. For the cutting of a plasma membrane during bacterial cytokinesis, a tubulin homolog FtsZ protein is recruited from the cytoplasm to the division site. FtsZ protein polymerizes in a GTP-dependent manner and its N-terminal domain has a GTPase activity. In this study, we have begun to characterize FtsZ from Staphylococcus aureus (SA). Full-length SA FtsZ was cloned into pRSFDuet-1 vector and the clone was transformed into a BL21 (DE3) star cell. The recombinant SA FtsZ protein was purified using Ni-NTA affinity chromatography and dialysis. Using a spectrofluorometer, we showed that SA FtsZ undergoes a GTP-dependant polymerization in vitro. The polymer of the SA FtsZ protein disappeared after a few minutes, suggesting that the polymer is degraded as the GTP is consumed. This assay system may well be applied for inhibitor screening targeting S. aureus FtsZ.
Keywords
Staphylococcus aureus; FtsZ; polymerization assay; GTPase;
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1 Adams, D. W. and J. Errington. 2009. Bacterial cell division: assembly, maintenance and disassembly of the Z ring. Nat. Rev. Microbiol. 7: 642-653.   DOI   ScienceOn
2 Andreu J. M., C. Schaffner-Barbero, S. Huecas, D. Alonso, M. L. Lopez-Rodriguez, L. B. Ruiz-Avila, R. Nunez-Ramirez, O. Llorca, and A. J. Martin-Galiano. 2010. The antibacterial cell division inhibitor PC190723 is an FtsZ polymer-stabilizing agent that induces filament assembly and condensation. J. Biol. Chem. 285: 14239-14246.   DOI
3 Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.   DOI   ScienceOn
4 de Boer, P., R. Crossley, and L. Rothfield. 1992. The essential bacterial cell-division protein FtsZ is a GTPase. Nature 359: 254-256.   DOI   ScienceOn
5 Haydon, D. J., N. R. Stokes, R. Ure, G. Galbraith, J. M. Bennett, D. R. Brown, P. J. Baker, V. V. Barynin, D. W. Rice, S. E. Sedelnikova, J. R. Heal, J. M. Sheridan, S. T. Aiwale, P. K. Chauhan, A. Srivastava, A. Taneja, I. Collins, J. Errington, and L. G. Czaplewski. 2008. An inhibitor of FtsZ with potent and selective anti-staphylococcal activity. Science 321: 1673-1675.   DOI
6 Jevons, M. P., A. W. Coe, and M. T. Parker. 1963. Methicillin resistance in Staphylococci. Lancet 1: 904-907.
7 Kwon, Y. I., T. W. Kim, H. Y. Kim, Y. H. Chang, H. S. Kwak, G. J. Woo, and Y. H. Chung. 2007. Monitoring of methicillin resistant Staphylococcus aureus from medical environment in Korea. Kor. J. Microbiol. Biotechnol. 35: 158-162.
8 Lowe, J. and L. A. Amos. 1998. Crystal structure of the bacterial cell-division protein FtsZ. Nature 391: 203-206.   DOI   ScienceOn
9 Mukherjee, A. and J. Lutkenhaus. 1998. Dynamic assembly of FtsZ regulated by GTP hydrolysis. EMBO J 17: 462-469.   DOI   ScienceOn
10 Mukherjee, A. and J. Lutkenhaus. 1999. Analysis of FtsZ assembly by light scattering and determination of the role of divalent metal cations. J. Bacteriol. 181: 823-832.
11 Osawa, M., D. E. Anderson, and H. P. Erickson. 2009. Curved FtsZ protolaments generate bending forces on liposome membranes. EMBO J 28: 3476-3484.   DOI   ScienceOn