The structure of Sulfolobus solfataricus 2-keto-3-deoxygluconate kinase

Acta Crystallogr D Biol Crystallogr. 2008 Dec;64(Pt 12):1283-7. doi: 10.1107/S0907444908036111. Epub 2008 Nov 18.

Abstract

The hyperthermophilic archaeon Sulfolobus solfataricus grows optimally above 353 K and utilizes an unusual promiscuous nonphosphorylative Entner-Doudoroff pathway to metabolize both glucose and galactose. It has been proposed that a part-phosphorylative Entner-Doudoroff pathway occurs in parallel in S. solfataricus, in which the 2-keto-3-deoxygluconate kinase (KDGK) is promiscuous for both glucose and galactose metabolism. Recombinant S. solfataricus KDGK protein was expressed in Escherichia coli, purified and crystallized in 0.1 M sodium acetate pH 4.1 and 1.4 M NaCl. The crystal structure of apo S. solfataricus KDGK was solved by molecular replacement to a resolution of 2.0 A and a ternary complex with 2-keto-3-deoxygluconate (KDGlu) and an ATP analogue was resolved at 2.1 A. The complex suggests that the structural basis for the enzyme's ability to phosphorylate KDGlu and 2-keto-3-deoxygalactonate (KDGal) is derived from a subtle repositioning of residues that are conserved in homologous nonpromiscuous kinases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives
  • Adenosine Triphosphate / chemistry
  • Adenosine Triphosphate / metabolism*
  • Crystallization
  • Enzyme Activation
  • Galactose / metabolism
  • Gluconates / metabolism*
  • Glucose / metabolism
  • Phosphorylation
  • Phosphotransferases (Alcohol Group Acceptor) / chemistry*
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Protein Binding
  • Protein Conformation
  • Structure-Activity Relationship
  • Substrate Specificity
  • Sulfolobus solfataricus / enzymology*
  • Ternary Complex Factors / chemistry
  • Ternary Complex Factors / metabolism*

Substances

  • Gluconates
  • Ternary Complex Factors
  • 2-keto-3-deoxygluconate
  • Adenosine Triphosphate
  • Phosphotransferases (Alcohol Group Acceptor)
  • 2-keto-3-deoxygluconokinase
  • Glucose
  • Galactose