Crystallization and preliminary X-ray crystallographic analysis of tRNA(m1G37)methyltransferase from Haemophilus influenzae

Acta Crystallogr D Biol Crystallogr. 2003 Jan;59(Pt 1):183-4. doi: 10.1107/s0907444902019716. Epub 2002 Dec 19.

Abstract

The enzyme tRNA(m(1)G37)methyltransferase (TrmD) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) specifically to guanosine at position 37 within a subset of tRNA species in bacteria. The modified guanosine is next to the anticodon and is important for the maintenance of the correct reading frame during translation. TrmD from Haemophilus influenzae with both N- and C-terminal tags was overexpressed in Escherichia coli and crystallized at 297 K using sodium acetate as a precipitant. Native X-ray diffraction data were collected to 1.85 A resolution. The crystals are rhombohedral, belonging to the space group R32, with unit-cell parameters a = b = 98.05, c = 176.79 A, alpha = beta = 90, gamma = 120 degrees. The presence of one monomer of recombinant TrmD in the crystallographic asymmetric unit gives a V(M) of 3.07 A(3) Da(-1) and a solvent content of 59.9%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Crystallization
  • Crystallography, X-Ray
  • DNA Primers / genetics
  • Escherichia coli / metabolism
  • Haemophilus influenzae / enzymology*
  • Molecular Sequence Data
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • tRNA Methyltransferases / chemistry*
  • tRNA Methyltransferases / genetics
  • tRNA Methyltransferases / metabolism

Substances

  • DNA Primers
  • Recombinant Proteins
  • tRNA Methyltransferases
  • tRNA (guanine-N1-)-methyltransferase