<> "The repository administrator has not yet configured an RDF license."^^ . <> . . . "Equilibrium binding and kinetic characterization of putative tetracycline repressor family transcription regulator Fad35R from Mycobacterium tuberculosis."^^ . "Fatty acids play critical role in the survival and virulence of Mycobacterium tuberculosis (Mtb). Activation of fatty acids by acyl-CoA synthetases (Fad) into fatty acyl-CoA is the first and one of the crucial steps in fatty acid metabolism. Mtb possesses 36 fatty acyl-CoA synthetases, unlike Escherichia coli, which has single enzyme. However, the mechanisms by which the expression of these multiple Fad genes is regulated remain uncharacterized. We characterized the DNA- and ligand-binding properties of a putative tetracycline repressor family regulator, named Fad35R, located upstream of the Fad35 gene and ScoA-citE operon. We identified a palindromic regulatory motif upstream of Fad35 and characterized the binding of Fad35R to this motif. Equilibrium binding studies show that Fad35R binds to this motif with high affinity (K(d) ∼ 0.033 μm) and the specificity of binding was confirmed by an electromobility gel shift assay. Kinetic studies indicate that faster association (k(a,avg) ∼ 5.4 × 10(4) m(-1) · s(-1)) and slower dissociation rates (k(d,avg) ∼ 5.84 × 10(-4) s(-1)) confer higher affinity. The affinity for the promoter is maximum at 300 mm NaCl but decreases rapidly beyond this range. Ligand-binding studies indicate that Fad35R binds specifically to tetracycline and also binds to fatty acid derivatives. The promoter-binding affinity is decreased significantly in the presence of palmityl-CoA, suggesting that Fad35R can sense the levels of activated fatty acids and alter its DNA-binding activity. Our results suggest that Fad35R may be the functional homologue of FadR and controls the expression of genes in a metabolite-dependent manner."^^ . "2012-09" . . "279" . "17" . . "John and Wiley"^^ . . . "The FEBS journal"^^ . . . "17424658" . . . . . . . . . . . . . . . . . . . . . . . . . "Sangaralingam"^^ . "Kumaran"^^ . "Sangaralingam Kumaran"^^ . . "Appu Kumar"^^ . "Singh"^^ . "Appu Kumar Singh"^^ . . "Bala"^^ . "Subramani"^^ . "Bala Subramani"^^ . . "Sushma"^^ . "Anand"^^ . "Sushma Anand"^^ . . "Vijay"^^ . "Singh"^^ . "Vijay Singh"^^ . . "Manish"^^ . "Datt"^^ . "Manish Datt"^^ . . "Monica"^^ . "Mittal"^^ . "Monica Mittal"^^ . . . . . . "Equilibrium binding and kinetic characterization of putative tetracycline repressor family transcription regulator Fad35R from Mycobacterium tuberculosis. (PDF)"^^ . . . . "HTML Summary of #1307 \n\nEquilibrium binding and kinetic characterization of putative tetracycline repressor family transcription regulator Fad35R from Mycobacterium tuberculosis.\n\n" . "text/html" . . . "QR Microbiology"@en . .