Expression purification crystallization and preliminary X-ray analysis of Pseudomonas aeruginosa PelD
The production of the PEL polysaccharide in Pseudomonas aeruginosa requires the binding of bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP) to the cytoplasmic GGDEF domain of the inner membrane protein PelD. Here, the overexpression, purification and crystallization of a soluble construct of PelD that encompasses the GGDEF domain and a predicted GAF domain is reported. Diffraction-quality crystals were grown using the hanging-drop vapour-diffusion method. The crystals grew as flat plates, with unit-cell parameters a = 88.3, b = 114.0, c = 61.9 {angstrom}, {alpha} = {beta} = {gamma} = 90.0{sup o}. The PelD crystals exhibited the symmetry of space group P2{sub 1}2{sub 1}2 and diffracted to a minimum d-spacing of 2.2 {angstrom}. On the basis of the Matthews coefficient (V{sub M} = 2.29 {angstrom}{sup 3} Da{sup -1}), it was estimated that two molecules are present in the asymmetric unit.
- Research Organization:
- Brookhaven National Lab. (BNL), Upton, NY (United States)
- Sponsoring Organization:
- USDOE SC OFFICE OF BIOLOGICAL & ENVIRONMENTAL RESEARCH
- DOE Contract Number:
- DE-AC02-98CH10886
- OSTI ID:
- 1041627
- Report Number(s):
- BNL-96931-2012-JA; R&D Project: BO-070; KP1605010; TRN: US201212%%45
- Journal Information:
- Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 68, Issue 2; ISSN 1744-3091
- Country of Publication:
- United States
- Language:
- English
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