Open AccessCrystallization and preliminary crystallographic analysis of tRNA (m 7 G46) methyltransferase from Escherichia coli
Author(s) -
Liu Qi,
Gao Yang,
Yang Weili,
Zhou Huihao,
Gao Yongxiang,
Zhang Xiao,
Teng Maikun,
Niu Liwen
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108020241
Subject(s) - transfer rna , escherichia coli , polyethylene glycol , methyltransferase , crystallography , crystallization , chemistry , peg ratio , methionine , crystal structure , stereochemistry , rna , methylation , amino acid , biochemistry , dna , organic chemistry , finance , economics , gene
Transfer RNA (tRNA) (m 7 G46) methyltransferase (TrmB) belongs to the Rossmann‐fold methyltransferase (RFM) family and uses S ‐adenosyl‐ l ‐methionine (SAM) as the methyl‐group donor to catalyze the formation of N 7 ‐methylguanosine (m 7 G) at position 46 in the variable loop of tRNAs. After attempts to crystallize full‐length Escherichia coli TrmB (EcTrmB) failed, a truncated protein lacking the first 32 residues of the N‐terminus but with an additional His 6 tag at the C‐terminus was crystallized by the hanging‐drop vapour‐diffusion method using polyethylene glycol 3350 (PEG 3350) as precipitant at 283 K. An X‐ray diffraction data set was collected using a single flash‐cooled crystal that belonged to space group P 2 1 .