PMID:12454498
| Citation |
Wright, A, Blewett, A, Fulop, V, Cooper, R, Burrows, S, Jones, C and Roper, D (2002) Expression, purification, crystallization and preliminary characterization of an HHED aldolase homologue from Escherichia coli K12. Acta Crystallogr. D Biol. Crystallogr. 58:2191-3 |
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| Abstract |
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 A 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 A. A 90% complete data set was collected to 1.78 A from a single native crystal using in-house facilities. |
| Links | |
| Keywords |
Aldehyde-Lyases/chemistry; Aldehyde-Lyases/genetics; Aldehyde-Lyases/isolation & purification; Cloning, Molecular; Crystallization; Crystallography, X-Ray; Escherichia coli/enzymology; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/isolation & purification; Protein Conformation |
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