Crystallization and preliminary X-ray analysis of the tRNA thiolation enzyme MnmA from Escherichia coli complexed with tRNA super(Glu)

MnmA catalyzes a sulfuration reaction to synthesize 2-thiouridine at the wobble positions of tRNA super(Glu), tRNA super(Gln) and tRNA super(Lys) in Escherichia coli. The binary complex of MnmA and tRNA super(Glu) was crystallized in two different crystal forms: forms I and II. Cocrystallization of MnmA-tRNA super(Glu) with ATP yielded form III crystals. The three crystal forms diffracted to 3.1, 3.4 and 3.4 Aa resolution, respectively, using synchrotron radiation at SPring-8. These crystals belong to space groups C2, I2 sub(1)2 sub(1)2 sub(1) and C2, with unit-cell parameters a = 225.4, b = 175.8, c = 53.0 Aa, beta = 101.6 degree , a = 101.5, b = 108.0, c = 211.2 Aa and a = 238.1, b = 102.1, c = 108.2 Aa, beta = 117.0 degree , respectively. The asymmetric units of these crystals are expected to contain two, one and two MnmA-tRNA super(Glu) complexes, respectively.