Molecules and Cells

  • 제19권3호
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  • Pages.398-401
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  • 2005
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  • 1016-8478(pISSN)
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  • 0219-1032(eISSN)
한국분자세포생물학회 (Korean Society for Molecular and Cellular Biology)
권호 표지

Crystallization and Preliminary X-Ray Crystallographic Analysis of UDP-N-Acetylglucosamine Enolpyruvyl Transferase from Haemophilus influenzae in Complex with UDP-N-Acetylglucosamine and Fosfomycin

  • Yoon, Hye-Jin (Department of Chemistry, College of Natural Sciences, Seoul National University) ;
  • Ku, Min-Je (Department of Chemistry, College of Natural Sciences, Seoul National University) ;
  • Ahn, Hyung Jun (Department of Chemistry, College of Natural Sciences, Seoul National University) ;
  • Lee, Byung Il (Department of Chemistry, College of Natural Sciences, Seoul National University) ;
  • Mikami, Bunzo (Laboratory of Quality Design and Exploitation, Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University) ;
  • Suh, Se Won (Department of Chemistry, College of Natural Sciences, Seoul National University)
  • 투고 : 2005.01.21
  • 심사 : 2005.02.23
  • 발행 : 2005.06.30
⟨ 이전 논문 다음 논문 ⟩

초록

The bacterial enzyme UDP-N-acetylglucosamine enolpyruvyl transferase catalyzes the first committed step of peptidoglycan biosynthesis, i.e., transfer of enolpyruvate from phosphoenolpyruvate to UDP-N-acetyl-glucosamine. We have overexpressed the enzyme from Haemophilus influenzae in Escherichia coli and crystallized it in the apo-form, as well as in a complex with UDP-N-acetylglucosamine and fosfomycin using ammonium sulfate as the precipitant. X-ray diffraction data from a crystal of the apo-form were collected to $2.8{\AA}$ resolution at 293 K. The crystal quality was improved by co-crystallization with UDP-N-acetylglucosamine and fosfomycin. X-ray data to $2.2{\AA}$ have been collected at 100 K from a flash-frozen crystal of the complex. The complex crystals belong to the orthorhombic space group I222 (or $I2_12_12_1$) with unit-cell parameters of a = 63.7, b = 124.5, and $c=126.3{\AA}$. Assuming a monomer of the recombinant enzyme in the crystallographic asymmetric unit, the calculated Matthews parameter ($V_M$) is $2.71{\AA}^3Da^{-1}$ and solvent content is 54.6%.

키워드

과제정보

연구 과제 주관 기관 : Korea Ministry of Science and Technology, Korea Science and Engineering Foundation

참고문헌

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