CRYSTALLIZATION OF HISTIDYL-TRANSFER-RNA SYNTHETASE FROM ESCHERICHIA-COLI

Citation
C. Francklyn et al., CRYSTALLIZATION OF HISTIDYL-TRANSFER-RNA SYNTHETASE FROM ESCHERICHIA-COLI, Journal of Molecular Biology, 241(2), 1994, pp. 275-277
Citations number
18
Categorie Soggetti
Biology
ISSN journal
00222836
Volume
241
Issue
2
Year of publication
1994
Pages
275 - 277
Database
ISI
SICI code
0022-2836(1994)241:2<275:COHSFE>2.0.ZU;2-F
Abstract
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.