Overproduction, crystallization and preliminary X-ray crystallographic analysis of Escherichia coli tRNA N-6-threonylcarbamoyladenosine dehydratase

Abstract

Escherichia coli tRNA N-6-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 angstrom resolution under cryoconditions using synchrotron X-rays. The crystals belonged to space group P2(1), with unit-cell parameters a = 65.4, b = 96.8, c = 83.3 angstrom, beta = 111.7 degrees. 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 (V-M) of 2.12 angstrom(3) Da(-1) and 42.1% solvent content.