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Crystallization and preliminary X-ray diffraction characterization of RpfF, a key DSF synthase from Stenotrophomonas

Xian Ya Lin1, Shu Ju Liao, Ko Hsin Chin

  • 1Institute of Biochemistry, National Chung-Hsing University, Taichung 40227, Taiwan.

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
|October 24, 2009
PubMed

Insights

Stenotrophomonas maltophilia is a resistant nosocomial pathogen. Targeting its RpfF enzyme, crucial for virulence, offers a new therapeutic strategy, as demonstrated by successful protein crystallization.

Area of Science:

  • Microbiology
  • Structural Biology
  • Drug Discovery

Background:

  • Stenotrophomonas maltophilia is an opportunistic pathogen causing significant hospital-acquired infections.
  • Antibiotic resistance in S. maltophilia necessitates novel therapeutic approaches.
  • Virulence is regulated by diffusible signal factor (DSF) quorum sensing, involving the RpfF enzyme.

Purpose of the Study:

  • To investigate the potential of inhibiting S. maltophilia RpfF (SmRpfF) as a novel anti-virulence strategy.
  • To facilitate structure-based drug design by obtaining crystalline SmRpfF.

Main Methods:

  • Overexpression of SmRpfF in Escherichia coli.
  • Purification of the recombinant SmRpfF protein.
  • Crystallization of SmRpfF using the hanging-drop vapour-diffusion method.

Main Results:

  • SmRpfF was successfully overexpressed and purified.
  • Crystals of SmRpfF were obtained.
  • The crystals belonged to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2, diffracting to 2.25 A resolution.

Conclusions:

  • Inhibiting SmRpfF function is a promising strategy to combat S. maltophilia infections.
  • The obtained crystal structure provides a foundation for developing small-molecule inhibitors targeting SmRpfF.

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