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Title: Crystallization and avoiding the problem of hemihedral twinning in crystals of Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus

Abstract

Δ{sup 1}-Pyrroline-5-carboxylate dehydrogenase from T. thermophilus has been crystallized in a citrate-bound form. The crystals exhibited hemihedral twinning and untwinned crystals were obtained by exclusion of lithium sulfate from the crystallization solution. Apo, NAD{sup +} bound-, NADH bound- and glutamate-bound forms were also obtained and characterized by X-ray diffraction. Δ{sup 1}-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 Å 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 Å, and diffract to 1.08 Å. Crystals soaked in solution with NAD{sup +} (TtP5CDh-NAD), NADH (TtP5CDh-NADH) and glutamate (TtP5CDh-Glu) were also prepared and characterized.

Authors:
; ; ;  [1]
  1. Advanced Protein Crystallography Research Group, RIKEN Harima Institute, 1-1-1 Kouto, Mikazuki-cho, Sayo-gun, Hyogo 679-5148 (Japan)
Publication Date:
OSTI Identifier:
22356022
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 61; Journal Issue: Pt 6; Other Information: PMCID: PMC1952327; PMID: 16511109; PUBLISHER-ID: gx5062; OAI: oai:pubmedcentral.nih.gov:1952327; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CITRATES; CRYSTALLIZATION; CRYSTALS; MATHEMATICAL SOLUTIONS; RESOLUTION; SOLUTIONS; SPACE GROUPS; TWINNING; VARIATIONS; X-RAY DIFFRACTION

Citation Formats

Inagaki, Eiji, Takahashi, Hitomi, Kuroishi, Chizu, and Tahirov, Tahir H., E-mail: tahir@spring8.or.jp. Crystallization and avoiding the problem of hemihedral twinning in crystals of Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus. United Kingdom: N. p., 2005. Web. doi:10.1107/S1744309105016118.
Inagaki, Eiji, Takahashi, Hitomi, Kuroishi, Chizu, & Tahirov, Tahir H., E-mail: tahir@spring8.or.jp. Crystallization and avoiding the problem of hemihedral twinning in crystals of Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus. United Kingdom. doi:10.1107/S1744309105016118.
Inagaki, Eiji, Takahashi, Hitomi, Kuroishi, Chizu, and Tahirov, Tahir H., E-mail: tahir@spring8.or.jp. Wed . "Crystallization and avoiding the problem of hemihedral twinning in crystals of Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus". United Kingdom. doi:10.1107/S1744309105016118.
@article{osti_22356022,
title = {Crystallization and avoiding the problem of hemihedral twinning in crystals of Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus},
author = {Inagaki, Eiji and Takahashi, Hitomi and Kuroishi, Chizu and Tahirov, Tahir H., E-mail: tahir@spring8.or.jp},
abstractNote = {Δ{sup 1}-Pyrroline-5-carboxylate dehydrogenase from T. thermophilus has been crystallized in a citrate-bound form. The crystals exhibited hemihedral twinning and untwinned crystals were obtained by exclusion of lithium sulfate from the crystallization solution. Apo, NAD{sup +} bound-, NADH bound- and glutamate-bound forms were also obtained and characterized by X-ray diffraction. Δ{sup 1}-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 Å 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 Å, and diffract to 1.08 Å. Crystals soaked in solution with NAD{sup +} (TtP5CDh-NAD), NADH (TtP5CDh-NADH) and glutamate (TtP5CDh-Glu) were also prepared and characterized.},
doi = {10.1107/S1744309105016118},
journal = {Acta Crystallographica. Section F},
number = Pt 6,
volume = 61,
place = {United Kingdom},
year = {Wed Jun 01 00:00:00 EDT 2005},
month = {Wed Jun 01 00:00:00 EDT 2005}
}