Overproduction, crystallization and preliminary X-ray crystallographic analysis of Escherichia coli tRNA N6-threonylcarbamoyladenosine dehydratase

Acta Crystallogr F Struct Biol Commun. 2014 Nov;70(Pt 11):1517-20. doi: 10.1107/S2053230X14020883. Epub 2014 Oct 25.

Abstract

Escherichia coli tRNA N6-threonylcarbamoyladenosine dehydratase (TcdA), previously called CsdL or YgdL, was overproduced and purified from E. coli and crystallized using polyethylene glycol 3350 as a crystallizing agent. X-ray diffraction data were collected to 2.70 Å resolution under cryoconditions using synchrotron X-rays. The crystals belonged to space group P2₁, with unit-cell parameters a=65.4, b=96.8, c=83.3 Å, β=111.7°. According to the Matthews coefficient, the asymmetric unit may contain up to four subunits of the monomeric protein, with a crystal volume per protein mass (VM) of 2.12 Å3 Da(-1) and 42.1% solvent content.

Keywords: CsdL; N6-threonylcarbamoyladenosine dehydratase; TcdA; tRNA hypermodification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / biosynthesis
  • Adenosine / chemistry
  • Adenosine / genetics
  • Amino Acid Sequence
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli Proteins / biosynthesis
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / genetics
  • Molecular Sequence Data
  • RNA, Transfer / biosynthesis
  • RNA, Transfer / chemistry*
  • RNA, Transfer / genetics
  • Threonine Dehydratase / biosynthesis
  • Threonine Dehydratase / chemistry*
  • Threonine Dehydratase / genetics

Substances

  • Escherichia coli Proteins
  • N(6)-(N-threonylcarbonyl)adenosine
  • RNA, Transfer
  • Threonine Dehydratase
  • Adenosine