Purification, crystallization and preliminary crystallographic analysis of the 23S rRNA methyltransferase RlmM (Cm2498) from Escherichia coli

RlmM is an AdoMet‐dependent methyltransferase that is responsible for 2′‐O‐methylation of C2498 in the peptidyl‐transferase loop of bacterial 23S rRNA. This modification occurs before assembly of the 50S ribosomal subunit, and lack of C2498 methylation can cause a slight reduction in bacterial fitne...

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Published in:Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2013-06, Vol.69 (6), p.640-642
Main Authors: Guo, Hong-Yue, Gao, Zeng-Qiang, Zhang, Heng, Wei, Yong, Xu, Jian-Hua, Wang, Wen-Ya, Yan, Ai-xia, Dong, Yu-Hui
Format: Article
Language:English
Subjects:
AdoMet
Crystallization Communications
Crystallography, X-Ray
Escherichia coli
Escherichia coli Proteins - chemistry
Escherichia coli Proteins - isolation & purification
methyltransferases
Methyltransferases - chemistry
Methyltransferases - isolation & purification
RlmM
RNA modification
RNA, Ribosomal, 23S - chemistry
RNA, Ribosomal, 23S - isolation & purification
X-Ray Diffraction
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Summary:RlmM is an AdoMet‐dependent methyltransferase that is responsible for 2′‐O‐methylation of C2498 in the peptidyl‐transferase loop of bacterial 23S rRNA. This modification occurs before assembly of the 50S ribosomal subunit, and lack of C2498 methylation can cause a slight reduction in bacterial fitness. Here, the purification and crystallization of RlmM from Escherichia coli as well as its preliminary crystallographic analysis are presented. Cocrystallization of RlmM with AdoMet was carried out and X‐ray diffraction data were collected to a resolution of 2.30 Å on beamline BL17U at the SSRF. However, electron density for AdoMet cannot be observed by comprehensive crystallographic analysis, indicating that it is not bound by RlmM during the cocrystallization process. The structure was solved by molecular replacement and refinement is in progress. The crystal contained one molecule in the asymmetric unit and belonged to space group P21, with unit‐cell parameters a = 56.07, b = 59.38, c = 54.35 Å, β = 94.84°, which differs from the P31 or P3121 space groups of previously reported RlmM structures (PDB entries 4auk, 4atn and 4b17).
ISSN:1744-3091
1744-3091
2053-230X
DOI:10.1107/S1744309113006611