3o74

Crystal structure of Cra transcriptional dual regulator from Pseudomonas putida

spinqualitylabelsall models 3o74, resolution 2.00Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 3o74 is a 2 chain structure with sequence from Pseudomonas putida KT2440. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2Å Ligands: GOL, MSE Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function Q88PQ6_PSEPK Publication Abstract from PubMed The catabolite repressor/activator (Cra) protein is a global sensor and regulator of carbon fluxes through the central metabolic pathways of gram-negative bacteria. To examine the nature of the effector (or effectors) that signal such fluxes to the protein of Pseudomonas putida, the Cra factor of this soil microorganism has been purified and characterized and its three-dimensional structure determined. Analytical ultracentrifugation, gel filtration, and mobility shift assays showed that the effector-free Cra is a dimer that binds an operator DNA sequence in the promoter region of the fruBKA cluster. Furthermore, fructose 1-phosphate (F1P) was found to most efficiently dissociate the Cra-DNA complex. Thermodynamic parameters of the F1P-Cra-DNA interaction calculated by isothermal titration calorimetry revealed that the factor associates tightly to the DNA sequence 5'-TTAAACGTTTCA-3' (K(D) = 26.3 +/- 3.1 nM) and that F1P binds the protein with an apparent stoichiometry of 1.06 +/- 0.06 molecules per Cra monomer and a K(D) of 209 +/- 20 nM. Other possible effectors, like fructose 1,6-bisphosphate, did not display a significant affinity for the regulator under the assay conditions. Moreover, the structure of Cra and its co-crystal with F1P at a 2-A resolution revealed that F1P fits optimally the geometry of the effector pocket. Our results thus single out F1P as the preferred metabolic effector of the Cra protein of P. putida. Fructose 1-phosphate is the preferred effector of the metabolic regulator Cra of Pseudomonas putida.,Chavarria M, Santiago C, Platero R, Krell T, Casasnovas JM, de Lorenzo V J Biol Chem. 2011 Mar 18;286(11):9351-9. Epub 2011 Jan 14. PMID:21239488 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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