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Purification, crystallization and preliminary X-ray crystallographic analysis of 23S RNA m(2)G2445 methyltransferase RlmL from Escherichia coli

Wang, Kai Tuo; Ma, Linglong; Nan, Jie LU ; Su, Xiao Dong LU and Li, Lanfen (2010) In Acta Crystallographica. Section F: Structural Biology and Crystallization Communications2005-01-01+01:002014-01-01+01:00 66(Pt 11). p.6-1484
Abstract

The RlmL (YcbY) protein in Escherichia coli is an rRNA methyltransferase that is specific for m(2)G2445 modification of 23S RNA. The rlmL gene was cloned into the expression vector pET28a and expressed in the host E. coli strain BL21 (DE3). Recombinant protein with a six-histidine tag was purified by Ni(2+)-affinity chromatography followed by gel filtration. Crystals were grown using the hanging-drop vapour-diffusion method and a detergent was used as an additive to improve diffraction quality. The final crystals diffracted to 2.2 Å resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.6, b = 140.8, c = 102.9 Å, β = 102.3°. The crystal has a most probable solvent content of 62.8% with two molecules in... (More)

The RlmL (YcbY) protein in Escherichia coli is an rRNA methyltransferase that is specific for m(2)G2445 modification of 23S RNA. The rlmL gene was cloned into the expression vector pET28a and expressed in the host E. coli strain BL21 (DE3). Recombinant protein with a six-histidine tag was purified by Ni(2+)-affinity chromatography followed by gel filtration. Crystals were grown using the hanging-drop vapour-diffusion method and a detergent was used as an additive to improve diffraction quality. The final crystals diffracted to 2.2 Å resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.6, b = 140.8, c = 102.9 Å, β = 102.3°. The crystal has a most probable solvent content of 62.8% with two molecules in the asymmetric unit.

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author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Crystallization, Crystallography, X-Ray, Escherichia coli, Escherichia coli Proteins, Methyltransferases
in
Acta Crystallographica. Section F: Structural Biology and Crystallization Communications2005-01-01+01:002014-01-01+01:00
volume
66
issue
Pt 11
pages
3 pages
publisher
Wiley-Blackwell
external identifiers
  • scopus:78149311116
ISSN
2053-230X
DOI
language
English
LU publication?
yes
id
9a17072e-0bd3-49e4-8f3e-99f229f6bc84
date added to LUP
2016-09-07 22:51:37
date last changed
2018-05-29 12:02:40
@article{9a17072e-0bd3-49e4-8f3e-99f229f6bc84,
  abstract     = {<p>The RlmL (YcbY) protein in Escherichia coli is an rRNA methyltransferase that is specific for m(2)G2445 modification of 23S RNA. The rlmL gene was cloned into the expression vector pET28a and expressed in the host E. coli strain BL21 (DE3). Recombinant protein with a six-histidine tag was purified by Ni(2+)-affinity chromatography followed by gel filtration. Crystals were grown using the hanging-drop vapour-diffusion method and a detergent was used as an additive to improve diffraction quality. The final crystals diffracted to 2.2 Å resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.6, b = 140.8, c = 102.9 Å, β = 102.3°. The crystal has a most probable solvent content of 62.8% with two molecules in the asymmetric unit.</p>},
  author       = {Wang, Kai Tuo and Ma, Linglong and Nan, Jie and Su, Xiao Dong and Li, Lanfen},
  issn         = {2053-230X},
  keyword      = {Crystallization,Crystallography, X-Ray,Escherichia coli,Escherichia coli Proteins,Methyltransferases},
  language     = {eng},
  month        = {11},
  number       = {Pt 11},
  pages        = {6--1484},
  publisher    = {Wiley-Blackwell},
  series       = {Acta Crystallographica. Section F: Structural Biology and Crystallization Communications2005-01-01+01:002014-01-01+01:00},
  title        = {Purification, crystallization and preliminary X-ray crystallographic analysis of 23S RNA m(2)G2445 methyltransferase RlmL from Escherichia coli},
  url          = {http://dx.doi.org/},
  volume       = {66},
  year         = {2010},
}