Crystallization and preliminary X-ray crystallographic study of the editing domain of Thermus thermophilus isoleucyl-tRNA synthetase complexed with pre- and post-transfer editing-substrate analogues

Ryuya Fukunaga, Shigeyuki Yokoyama

Research output: Contribution to journalArticle

Abstract

The CP1 domain (the editing domain) of isoleucyl-tRNA synthetase (IleRS) hydrolyzes misactivated Val-AMP in pre-transfer editing and mischarged Val-tRNAIle in post-transfer editing. The CP1 domain of Thermus thermophilus IleRS was expressed in isolation, purified and cocrystallized with Val-AMS (a Val-AMP analogue) and with Val-2AA (a Val-tRNAIle analogue). Two different expression constructs were used for each cocrystallization. The complex crystals with Val-AMS belong to the tetragonal space group P41212, with unit-cell parameters a = b = 102.00, c = 84.88 Å. The asymmetric unit contains two molecules of the CP1 domain, with a corresponding crystal volume per protein weight of 2.7 Å3 Da-1 and a solvent content of 53.5%. The complex crystals with Val-2AA belong to the tetragonal space group P4122, with unit-cell parameters a = b = 72.59, c =83.68 Å. The asymmetric unit contains one molecule of the CP1 domain, with a corresponding crystal volume per protein weight of 2.8 Å3 Da-1 and a solvent content of 55.8%. Data sets diffracting to 1.7 Å resolution were collected from each single crystal at 100 K.

Original languageEnglish (US)
Pages (from-to)1900-1902
Number of pages3
JournalActa Crystallographica Section D: Biological Crystallography
Volume60
Issue number10
DOIs
StatePublished - Oct 2004
Externally publishedYes

Fingerprint

Isoleucine-tRNA Ligase
RNA, Transfer, Ile
Thermus thermophilus
editing
Adenosine Monophosphate
Crystallization
X-Rays
crystallization
analogs
Weights and Measures
X rays
Crystals
adenosine monophosphate
Substrates
Alpha Magnetic Spectrometer
Proteins
x rays
crystals
Molecules
proteins

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

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title = "Crystallization and preliminary X-ray crystallographic study of the editing domain of Thermus thermophilus isoleucyl-tRNA synthetase complexed with pre- and post-transfer editing-substrate analogues",
abstract = "The CP1 domain (the editing domain) of isoleucyl-tRNA synthetase (IleRS) hydrolyzes misactivated Val-AMP in pre-transfer editing and mischarged Val-tRNAIle in post-transfer editing. The CP1 domain of Thermus thermophilus IleRS was expressed in isolation, purified and cocrystallized with Val-AMS (a Val-AMP analogue) and with Val-2AA (a Val-tRNAIle analogue). Two different expression constructs were used for each cocrystallization. The complex crystals with Val-AMS belong to the tetragonal space group P41212, with unit-cell parameters a = b = 102.00, c = 84.88 {\AA}. The asymmetric unit contains two molecules of the CP1 domain, with a corresponding crystal volume per protein weight of 2.7 {\AA}3 Da-1 and a solvent content of 53.5{\%}. The complex crystals with Val-2AA belong to the tetragonal space group P4122, with unit-cell parameters a = b = 72.59, c =83.68 {\AA}. The asymmetric unit contains one molecule of the CP1 domain, with a corresponding crystal volume per protein weight of 2.8 {\AA}3 Da-1 and a solvent content of 55.8{\%}. Data sets diffracting to 1.7 {\AA} resolution were collected from each single crystal at 100 K.",
author = "Ryuya Fukunaga and Shigeyuki Yokoyama",
year = "2004",
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T1 - Crystallization and preliminary X-ray crystallographic study of the editing domain of Thermus thermophilus isoleucyl-tRNA synthetase complexed with pre- and post-transfer editing-substrate analogues

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N2 - The CP1 domain (the editing domain) of isoleucyl-tRNA synthetase (IleRS) hydrolyzes misactivated Val-AMP in pre-transfer editing and mischarged Val-tRNAIle in post-transfer editing. The CP1 domain of Thermus thermophilus IleRS was expressed in isolation, purified and cocrystallized with Val-AMS (a Val-AMP analogue) and with Val-2AA (a Val-tRNAIle analogue). Two different expression constructs were used for each cocrystallization. The complex crystals with Val-AMS belong to the tetragonal space group P41212, with unit-cell parameters a = b = 102.00, c = 84.88 Å. The asymmetric unit contains two molecules of the CP1 domain, with a corresponding crystal volume per protein weight of 2.7 Å3 Da-1 and a solvent content of 53.5%. The complex crystals with Val-2AA belong to the tetragonal space group P4122, with unit-cell parameters a = b = 72.59, c =83.68 Å. The asymmetric unit contains one molecule of the CP1 domain, with a corresponding crystal volume per protein weight of 2.8 Å3 Da-1 and a solvent content of 55.8%. Data sets diffracting to 1.7 Å resolution were collected from each single crystal at 100 K.

AB - The CP1 domain (the editing domain) of isoleucyl-tRNA synthetase (IleRS) hydrolyzes misactivated Val-AMP in pre-transfer editing and mischarged Val-tRNAIle in post-transfer editing. The CP1 domain of Thermus thermophilus IleRS was expressed in isolation, purified and cocrystallized with Val-AMS (a Val-AMP analogue) and with Val-2AA (a Val-tRNAIle analogue). Two different expression constructs were used for each cocrystallization. The complex crystals with Val-AMS belong to the tetragonal space group P41212, with unit-cell parameters a = b = 102.00, c = 84.88 Å. The asymmetric unit contains two molecules of the CP1 domain, with a corresponding crystal volume per protein weight of 2.7 Å3 Da-1 and a solvent content of 53.5%. The complex crystals with Val-2AA belong to the tetragonal space group P4122, with unit-cell parameters a = b = 72.59, c =83.68 Å. The asymmetric unit contains one molecule of the CP1 domain, with a corresponding crystal volume per protein weight of 2.8 Å3 Da-1 and a solvent content of 55.8%. Data sets diffracting to 1.7 Å resolution were collected from each single crystal at 100 K.

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