The quaternary structure of the amidase from Geobacillus pallidus RAPc8 is revealed by its crystal packing
Journal Article
2006
Permanent link to this Item
Authors
Journal Title
Acta Crystallographica. Section F
Link to Journal
Journal ISSN
Volume Title
Publisher
International Union of Crystallography
Publisher
University of Cape Town
License
Series
Abstract
The amidase from Geobacillus pallidus RAPc8, a moderate thermophile, is a member of the nitrilase enzyme superfamily. It converts amides to the corresponding acids and ammonia and has application as an industrial catalyst. RAPc8 amidase has been cloned and functionally expressed in Escherichia coli and has been purified by heat treatment and a number of chromatographic steps. The enzyme was crystallized using the hanging-drop vapour-diffusion method. Crystals produced in the presence of 1.2 M sodium citrate, 400 mM NaCl, 100 mM sodium acetate pH 5.6 were selected for X-ray diffraction studies. A data set having acceptable statistics to 1.96 A˚ resolution was collected under cryoconditions using an in-house X-ray source. The space group was determined to be primitive cubic P4232, with unit-cell parameter a = 130.49 ( 0.05) A˚ . The structure was solved by molecular replacement using the backbone of the hypothetical protein PH0642 from Pyrococcus horikoshii (PDB code 1j31) with all non-identical side chains substituted with alanine as a probe. There is one subunit per asymmetric unit. The subunits are packed as trimers of dimers with D3 point-group symmetry around the threefold axis in such a way that the dimer interface seen in the homologues is preserved.
Description
Reference:
Agarkar, V. B., Kimani, S. W., Cowan, D. A., Sayed, M. R., & Sewell, B. T. (2006). The quaternary structure of the amidase from Geobacillus pallidus RAPc8 is revealed by its crystal packing. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 62(12), 1174-1178.