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 language | English (US) |
|---|---|
| Pages (from-to) | 1031-1033 |
| Number of pages | 3 |
| Journal | Acta Crystallographica Section F: Structural Biology and Crystallization Communications |
| Volume | 62 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2006 |
| Externally published | Yes |
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Keywords
- Methanosarcina mazei
- Pyrrolysyl-tRNA synthetase
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Structural Biology
- Genetics
- Condensed Matter Physics
Cite this
Crystallization and preliminary X-ray crystallographic analysis of the catalytic domain of pyrrolysyl-tRNA synthetase from the methanogenic archaeon Methanosarcina mazei. / Ishii, Ryohei; Nureki, Osamu; Yanagisawa, Tatsuo; Fukunaga, Ryuya; Yokoyama, Shigeyuki.
In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 62, No. 10, 10.2006, p. 1031-1033.Research output: Contribution to journal › Article
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TY - JOUR
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.
KW - Methanosarcina mazei
KW - Pyrrolysyl-tRNA synthetase
UR - http://www.scopus.com/inward/record.url?scp=33749524376&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33749524376&partnerID=8YFLogxK
U2 - 10.1107/S1744309106036700
DO - 10.1107/S1744309106036700
M3 - Article
VL - 62
SP - 1031
EP - 1033
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
SN - 1744-3091
IS - 10
ER -