Crystallization and avoiding the problem of hemihedral twinning in crystals of Delta1-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Jun 1;61(Pt 6):609-11. doi: 10.1107/S1744309105016118. Epub 2005 Jun 1.

Abstract

Delta1-Pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus (TtP5CDh) has been crystallized in a citrate-bound form (TtP5CDh-cit). The crystals diffracted to well beyond 2 A resolution, but exhibited perfect or near-perfect hemihedral twinning. Variation of crystallization conditions resulted in the growth of larger untwinned crystals or crystals with significantly reduced twin content, all with similar unit-cell parameters. The soaking of TtP5CDh-cit crystals in citrate-free solution produced crystals of the apo form (TtP5CDh-apo). The TtP5CDh-apo crystals belong to space group R3, with unit-cell parameters a = b = 102.29, c = 279.28 A, and diffract to 1.08 A. Crystals soaked in solution with NAD+ (TtP5CDh-NAD), NADH (TtP5CDh-NADH) and glutamate (TtP5CDh-Glu) were also prepared and characterized.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Pyrroline-5-Carboxylate Dehydrogenase / chemistry*
  • 1-Pyrroline-5-Carboxylate Dehydrogenase / genetics
  • 1-Pyrroline-5-Carboxylate Dehydrogenase / isolation & purification
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification
  • Citric Acid / chemistry
  • Crystallization / methods
  • Glutamic Acid / chemistry
  • NAD / chemistry
  • Polymerase Chain Reaction
  • Thermus thermophilus / enzymology*
  • X-Ray Diffraction

Substances

  • Bacterial Proteins
  • NAD
  • Citric Acid
  • Glutamic Acid
  • 1-Pyrroline-5-Carboxylate Dehydrogenase