Crystallization and avoiding the problem of hemihedral twinning in crystals of Delta1-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus.
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.
Inagaki
Eiji
E
Advanced Protein Crystallography Research Group, RIKEN Harima Institute, 1-1-1 Kouto, Sayo-gun, Hyogo 679-5148, Japan.
Takahashi
Hitomi
H
Kuroishi
Chizu
C
Tahirov
Tahir H
TH
D050842:1-Pyrroline-5-Carboxylate Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D019343:Citric Acid / Q000737:chemistry; D003460:Crystallization / Q000379:methods; D018698:Glutamic Acid / Q000737:chemistry; D009243:NAD / Q000737:chemistry; D016133:Polymerase Chain Reaction; D016963:Thermus thermophilus / Q000201:enzymology*; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D009243:NAD; D019343:Citric Acid; D018698:Glutamic Acid; D050842:1-Pyrroline-5-Carboxylate Dehydrogenase