Purification, crystallization and preliminary X-ray crystallographic analysis of 23S RNA m(2)G2445 methyltransferase RlmL from Escherichia coli
(2010) In Acta Crystallographica. Section F: Structural Biology and Crystallization Communications 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.
(Less)
- author
- Wang, Kai Tuo ; Ma, Linglong ; Nan, Jie LU ; Su, Xiao Dong LU and Li, Lanfen
- organization
- publishing date
- 2010-11-01
- 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 Communications
- volume
- 66
- issue
- Pt 11
- pages
- 3 pages
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:78149311116
- pmid:21045301
- ISSN
- 2053-230X
- DOI
- 10.1107/S1744309110035074
- language
- English
- LU publication?
- yes
- id
- 9a17072e-0bd3-49e4-8f3e-99f229f6bc84
- date added to LUP
- 2016-09-07 22:51:37
- date last changed
- 2024-04-05 05:58:55
@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}}, keywords = {{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 Communications}}, 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/10.1107/S1744309110035074}}, doi = {{10.1107/S1744309110035074}}, volume = {{66}}, year = {{2010}}, }