5hpi

Crystal Structure of the Double Mutant of PobR Transcription Factor Inducer Binding Domain-3-Hydroxy Benzoic Acid complex from AcinetobacterCrystal Structure of the Double Mutant of PobR Transcription Factor Inducer Binding Domain-3-Hydroxy Benzoic Acid complex from Acinetobacter

Structural highlights 5hpi is a 4 chain structure with sequence from Acinetobacter baylyi ADP1. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2.963Å Ligands: , , , Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function POBR_ACIAD Positive regulator of the pobA gene for p-hydroxybenzoate hydroxylase. Publication Abstract from PubMed A whole-cell biosensor utilizing a transcription factor (TF) is an effective tool for sensitive and selective detection of specialty chemicals or anthropogenic molecules, but requires access to an expanded repertoire of TFs. Using homology modeling and ligand docking for binding pocket identification, assisted by conservative mutations in the pocket, we engineered a novel specificity in an Acinetobacter TF, PobR, to 'sense' a chemical p-nitrophenol (pNP) and measured the response via a fluorescent protein reporter expressed from a PobR promoter. Out of 10(7) variants of PobR, four were active when dosed with pNP, with two mutants showing a specificity switch from the native effector 4-hydroxybenzoate (4HB). One of the mutants, pNPmut1 was then used to create a smart microbial cell responding to pNP production from hydrolysis of an insecticide, paraoxon, in a coupled assay involving phosphotriesterase (PTE) enzyme expressed from a separate promoter. We show the fluorescence of the cells correlated with the catalytic efficiency of the PTE variant expressed in each cell. High selectivity between similar molecules (4HB versus pNP), high sensitivity for pNP detection ( approximately 2 muM) and agreement of apo- and holo-structures of PobR scaffold with predetermined computational models are other significant results presented in this work. A microbial sensor for organophosphate hydrolysis exploiting an engineered specificity switch in a transcription factor.,Jha RK, Kern TL, Kim Y, Tesar C, Jedrzejczak R, Joachimiak A, Strauss CE Nucleic Acids Res. 2016 Sep 30;44(17):8490-500. doi: 10.1093/nar/gkw687. Epub, 2016 Aug 17. PMID:27536006 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

spinqualitylabelsall models 5hpi, resolution 2.96Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image