C. Francklyn et al., CRYSTALLIZATION OF HISTIDYL-TRANSFER-RNA SYNTHETASE FROM ESCHERICHIA-COLI, Journal of Molecular Biology, 241(2), 1994, pp. 275-277
Histidyl-tRNA synthetase from Escherichia coli was over-expressed and
purified by Q Sepharose and hydroxyapatite chromatography. Crystals of
the complex containing histidyl-tRNA synthetase, ATP and histidine ha
ve been grown by vapor diffusion against reservoirs containing 0.1 M T
ris (pH 7.4), 0.5 M NaCl and 10% polyethylene glycol 6000. Under these
conditions, two crystal forms are obtained. The triclinic form has un
it cell dimensions a = 61.3 Angstrom, b = 108.5 Angstrom, c = 110.2 An
gstrom, alpha = 115.1 degrees: beta = 90.2 degrees and gamma = 97.2 de
grees. The monoclinic form, space group P2(1), has cell dimensions a =
61.2 Angstrom, b = 109.7 Angstrom, c = 196.7 Angstrom and beta = 98.1
degrees. Both crystal forms diffract up to 2.7 Angstrom and are stabl
e in the synchrotron beam. Assuming a dimeric mass of 96,000 daltons a
nd V-m value of 3.4 Angstrom(3)/dalton, the asymmetric unit in both fo
rms contains two dimers with a solvent content of approximately 60%. A
3.7 Angstrom resolution native dataset with an R(merge) on intensitie
s of 7.9% has been collected from the monoclinic crystal form.