Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei

Ryohei Ishii, Osamu Nureki, Tatsuo Yanagisawa, Ryuya Fukunaga, Shigeyuki Yokoyama

Research output: Contribution to journalArticle

Abstract

Pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei was overexpressed in an N-terminally truncated form PylRS(c270) in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The native PylRS(c270) crystals in complex with an ATP analogue belonged to space group P64, with unit-cell parameters a = b = 104.88, c = 70.43 Å, α = β = 90, γ = 120°, and diffracted to 1.9 Å resolution. The asymmetric unit contains one molecule of PylRS(c270). Selenomethionine- substituted protein crystals were prepared in order to solve the structure by the MAD phasing method.

Original languageEnglish (US)
Pages (from-to)1031-1033
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume62
Issue number10
DOIs
StatePublished - Oct 2006
Externally publishedYes

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Methanosarcina
Amino Acyl-tRNA Synthetases
Archaea
Crystallization
Catalytic Domain
X-Rays
crystallization
X rays
adenosine triphosphate
Escherichia
crystals
homogeneity
glycols
polyethylenes
x rays
vapors
analogs
Selenomethionine
proteins
Crystals

Keywords

  • Methanosarcina mazei
  • Pyrrolysyl-tRNA synthetase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Structural Biology
  • Genetics
  • Condensed Matter Physics

Cite this

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title = "Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei",
abstract = "Pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei was overexpressed in an N-terminally truncated form PylRS(c270) in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The native PylRS(c270) crystals in complex with an ATP analogue belonged to space group P64, with unit-cell parameters a = b = 104.88, c = 70.43 {\AA}, α = β = 90, γ = 120°, and diffracted to 1.9 {\AA} resolution. The asymmetric unit contains one molecule of PylRS(c270). Selenomethionine- substituted protein crystals were prepared in order to solve the structure by the MAD phasing method.",
keywords = "Methanosarcina mazei, Pyrrolysyl-tRNA synthetase",
author = "Ryohei Ishii and Osamu Nureki and Tatsuo Yanagisawa and Ryuya Fukunaga and Shigeyuki Yokoyama",
year = "2006",
month = "10",
doi = "10.1107/S1744309106036700",
language = "English (US)",
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pages = "1031--1033",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
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T1 - Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei

AU - Ishii, Ryohei

AU - Nureki, Osamu

AU - Yanagisawa, Tatsuo

AU - Fukunaga, Ryuya

AU - Yokoyama, Shigeyuki

PY - 2006/10

Y1 - 2006/10

N2 - Pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei was overexpressed in an N-terminally truncated form PylRS(c270) in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The native PylRS(c270) crystals in complex with an ATP analogue belonged to space group P64, with unit-cell parameters a = b = 104.88, c = 70.43 Å, α = β = 90, γ = 120°, and diffracted to 1.9 Å resolution. The asymmetric unit contains one molecule of PylRS(c270). Selenomethionine- substituted protein crystals were prepared in order to solve the structure by the MAD phasing method.

AB - Pyrrolysyl-tRNA synthetase (PylRS) from Methanosarcina mazei was overexpressed in an N-terminally truncated form PylRS(c270) in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The native PylRS(c270) crystals in complex with an ATP analogue belonged to space group P64, with unit-cell parameters a = b = 104.88, c = 70.43 Å, α = β = 90, γ = 120°, and diffracted to 1.9 Å resolution. The asymmetric unit contains one molecule of PylRS(c270). Selenomethionine- substituted protein crystals were prepared in order to solve the structure by the MAD phasing method.

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