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 1 2006

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Keywords

  • Methanosarcina mazei
  • Pyrrolysyl-tRNA synthetase

ASJC Scopus subject areas

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

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