Expression, purification, crystallization and preliminary characterization of an HHED aldolase homologue from Escherichia coli K12
UNSPECIFIED. (2002) Expression, purification, crystallization and preliminary characterization of an HHED aldolase homologue from Escherichia coli K12. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 58 (Part 12). pp. 2191-2193. ISSN 0907-4449Full text not available from this repository.
Official URL: http://dx.doi.org/10.1107/S0907444902017894
An ORF designated b2245 (yfaU) in the Escherichia coli K12 genome sequence, identified as an HHED aldolase homologue, was cloned into the high-expression plasmid pT7-7 and overexpressed in E. coli B835(DE3). The enzyme was purified in three steps to 95% purity prior to crystallization. Crystals were obtained by the hanging-drop vapour-diffusion method at 277 K from a number of screening conditions. Crystals suitable for structural studies were grown from solutions containing 0.4 M ammonium dihydrogen phosphate and grew to a maximum dimension of approximately 0.5 mm. Diffraction data to 1.7 Angstrom were collected using an in-house Cu Kalpha radiation source at 100 K. The crystals belong to space group C222(1), with unit-cell parameters a = 105.1, b = 136.6, c = 123.1 Angstrom. A 90% complete data set was collected to 1.78 Angstrom from a single native crystal using in-house facilities.
|Item Type:||Journal Article|
|Subjects:||Q Science > QD Chemistry
Q Science > QH Natural history > QH301 Biology
|Journal or Publication Title:||ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY|
|Official Date:||December 2002|
|Number of Pages:||3|
|Page Range:||pp. 2191-2193|
Actions (login required)
Downloads per month over past year