Crystallization and preliminary crystallographic analysis of 2-keto-3-deoxygluconate kinase from Thermus thermophilus

Eiji Inagaki, Yoko Ukita, Maki Kumei, Yasuhiro Kajihara, Tahir H. Tahirov

Research output: Contribution to journalArticle

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Abstract

2-Keto-3-deoxygluconate kinase (KDGK) catalyzes the phosphorylation of 2-keto-3-deoxygluconate (KDG) to 2-keto-3-deoxy-6-phosphogluconate. Two crystal forms of KDGK from Thermus thermophilus were obtained by vapour-diffusion and microbatch methods. Crystals in the form of triangular plates (TtKDGK-1) were obtained that belong to space group P3, with unit-cell parameters a = b = 145.83, c = 74.63 Å, and diffract to 3.2 Å. These crystals exhibited nearly perfect hemihedral twinning. Assigning six subunits of TtKDGK to the asymmetric unit of the crystal corresponds to a 46.2% solvent content. A single plate-like crystal (TtKDGK-2) belonged to space group P63, with unit-cell parameters a = b = 84.83, c = 168.49 Å, and diffracts to 2.25 Å. This crystal exhibits only partial hemihedral twinning, with a twin fraction of 24.4%. Diffraction-quality crystals of TtKDGK with bound ATP (TtKDGK-ATP), a = b = 84.72, c = 321.61 Å and with bound KDG plus the ATP analogue AMP-PNP (TtKDGK-ATP-KDG), with unit-cell parameters a = b = 84.32, c = 168.7 Å, were also prepared and characterized.

Original languageEnglish (US)
Pages (from-to)761-763
Number of pages3
JournalActa Crystallographica Section D: Biological Crystallography
Volume60
Issue number4
DOIs
StatePublished - Apr 1 2004

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Thermus thermophilus
Crystallization
Adenosine Triphosphate
crystallization
Crystals
adenosine triphosphate
Adenylyl Imidodiphosphate
crystals
Twinning
twinning
Phosphorylation
cells
adenosine monophosphate
phosphorylation
2-keto-3-deoxygluconokinase
2-keto-3-deoxygluconate
Diffraction
Vapors
vapors
analogs

ASJC Scopus subject areas

  • Structural Biology

Cite this

Crystallization and preliminary crystallographic analysis of 2-keto-3-deoxygluconate kinase from Thermus thermophilus. / Inagaki, Eiji; Ukita, Yoko; Kumei, Maki; Kajihara, Yasuhiro; Tahirov, Tahir H.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 60, No. 4, 01.04.2004, p. 761-763.

Research output: Contribution to journalArticle

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abstract = "2-Keto-3-deoxygluconate kinase (KDGK) catalyzes the phosphorylation of 2-keto-3-deoxygluconate (KDG) to 2-keto-3-deoxy-6-phosphogluconate. Two crystal forms of KDGK from Thermus thermophilus were obtained by vapour-diffusion and microbatch methods. Crystals in the form of triangular plates (TtKDGK-1) were obtained that belong to space group P3, with unit-cell parameters a = b = 145.83, c = 74.63 {\AA}, and diffract to 3.2 {\AA}. These crystals exhibited nearly perfect hemihedral twinning. Assigning six subunits of TtKDGK to the asymmetric unit of the crystal corresponds to a 46.2{\%} solvent content. A single plate-like crystal (TtKDGK-2) belonged to space group P63, with unit-cell parameters a = b = 84.83, c = 168.49 {\AA}, and diffracts to 2.25 {\AA}. This crystal exhibits only partial hemihedral twinning, with a twin fraction of 24.4{\%}. Diffraction-quality crystals of TtKDGK with bound ATP (TtKDGK-ATP), a = b = 84.72, c = 321.61 {\AA} and with bound KDG plus the ATP analogue AMP-PNP (TtKDGK-ATP-KDG), with unit-cell parameters a = b = 84.32, c = 168.7 {\AA}, were also prepared and characterized.",
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