Crystallization and preliminary X-ray crystallographic characterization of TrmFO, a folate-dependent tRNA methyltransferase from Thermotoga maritima.
TrmFO, previously classified as GID, is a methyltransferase that catalyzes the formation of 5-methyluridine or ribothymidine (T) at position 54 in tRNA in some Gram-positive bacteria. To date, TrmFO is the only characterized tRNA methyltransferase that does not use S-adenosylmethionine as the methyl-group donor. Instead, the donor of the methyl group is N(5),N(10)-methylenetetrahydrofolate. The crystallization and preliminary X-ray crystallographic studies of TrmFO are reported here. The recombinant protein, cloned from Thermotoga maritima genomic DNA, was overproduced in Esherichia coli and crystallized in 25%(v/v) PEG 4000, 100 mM NaCl and sodium citrate buffer pH 5.0 at 291 K using the hanging-drop vapor-diffusion method. The plate-shaped crystals diffracted to 2.6 A and belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 79.94, b = 92.46, c = 127.20 A.
Cicmil
Nenad
N
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. cicmil@uiuc.edu
D003460:Crystallization; D018360:Crystallography, X-Ray; D020124:Thermotoga maritima / Q000201:enzymology* / Q000235:genetics; D012359:tRNA Methyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D012359:tRNA Methyltransferases