1991
DOI: 10.1016/s0021-9258(18)54258-7
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How does trp repressor bind to its operator?

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Cited by 53 publications
(13 citation statements)
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“…Under these conditions, TrpR is a dimer (K. S. Martin et al, submitted for publication). DNA-binding could not be measured by titration microcalorimetry, principally because of the magnitude of the dissociation constant, Ki (subnanomolar); the relative insensitivity of Ki to solution conditions (Carey, 1988;Carey et al, 1991) precluded shifting Ki into an accessible range. At TrpR and DNA concentrations far above Ki, multiple binding modes with different stoichiometries are used (Carey, 1988;Carey et al, 1991;Haran et al, 1992), complicating interpretation of calorimetric data.…”
Section: Methodsmentioning
confidence: 99%
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“…Under these conditions, TrpR is a dimer (K. S. Martin et al, submitted for publication). DNA-binding could not be measured by titration microcalorimetry, principally because of the magnitude of the dissociation constant, Ki (subnanomolar); the relative insensitivity of Ki to solution conditions (Carey, 1988;Carey et al, 1991) precluded shifting Ki into an accessible range. At TrpR and DNA concentrations far above Ki, multiple binding modes with different stoichiometries are used (Carey, 1988;Carey et al, 1991;Haran et al, 1992), complicating interpretation of calorimetric data.…”
Section: Methodsmentioning
confidence: 99%
“…DNase I Footprinting Titrations. Quantitative DNase I footprinting titration experiments (Brenowitz et al, 1986;Carey, 1988;Carey et al, 1991) were conducted to measure the affinity of TrpR for its operator DNA at different temperatures. The binding buffer for complex formation and TrpR dilution contained 2.5 mM sodium phosphate, pH 7.6, 25 mM NaCl, and 0.4 mM L-Trp.…”
Section: Methodsmentioning
confidence: 99%
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“…The presence of the corepressor L-tryptophan enhances the affinity of the repressor for the operator by initiating a conformational change to permit the interaction of the sequence-reading helices with two adjacent major grooves of the 18 base pair operator (Zhang et al, 1987). Solution assays including a phosphatase protection assay (Marmorstein et al, 1991) and gel retardation assays (Carey et al, 1991;Haran et al, 1992) both indicate high-affinity binding between the repressor and the operator sequence. However, the nature of the interactions at the interface between the protein and the nucleic acid as implicated by the crystal structures is unusual in that only a single direct amino acid-nucleobase contact appears to be present in each half-site: a bidentate interaction between Arg 69 and the base residue G -9 .…”
mentioning
confidence: 99%
“…When TR binds this site, it represses transcription of those genes whose protein products are responsible for the synthesis of tryptophan. The repressor protein has been extensively studied from a genetic as well as a structural standpoint (Kelley & Yanofsky, 1985 in inducing specific DNA binding is still not well understood (Carey et al, 1991;Fernando & Royer, 1992).…”
mentioning
confidence: 99%