Footprint titrations yield valid thermodynamic isotherms

M BRENOWITZ; D. F SENEAR; M. A SHEA; G. K ACKERS

doi:10.1073/pnas.83.22.8462

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Footprint titrations yield valid thermodynamic isotherms

Journal article

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Footprint titrations yield valid thermodynamic isotherms

M BRENOWITZ, D. F SENEAR, M. A SHEA and G. K ACKERS

Proceedings of the National Academy of Sciences - PNAS, Vol.83(22), pp.8462-8466

1986

DOI: 10.1073/pnas.83.22.8462

PMID: 3464963

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Abstract

A central issue in gene regulation is the mechanism, and biological function, of the cooperative binding of regulatory protein ligands to specific sites on DNA. To elucidate the physical-chemical basis of these interactions we have developed a thermodynamically rigorous method for conducting DNase I "footprint" (protection) titration experiments. The intrinsic binding constants and also those for cooperative interactions between various sites can be resolved from the individual-site binding curves determined by this technique. Experimental studies of cI-repressor-operator binding have demonstrated that the method provides an accurate representation of the fractional saturation of a binding site. We present individual-site binding curves for a lambda operator with two competent sites that demonstrate the presence of cooperative interactions between the sites. These curves set a lower limit to the magnitude of the cooperative free energy without comparison to single-site mutant operators.

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Intermolecular phenomena

Interactions. Associations

Molecular biophysics

Details

Title: Subtitle: Footprint titrations yield valid thermodynamic isotherms
Creators: M BRENOWITZ - Johns Hopkins univ., dep. biology, Baltimore MD 21218, United States
D. F SENEAR - Johns Hopkins univ., dep. biology, Baltimore MD 21218, United States
M. A SHEA - Johns Hopkins univ., dep. biology, Baltimore MD 21218, United States
G. K ACKERS - Johns Hopkins univ., dep. biology, Baltimore MD 21218, United States
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.83(22), pp.8462-8466
DOI: 10.1073/pnas.83.22.8462
PMID: 3464963
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; Washington, DC
Language: English
Date published: 1986
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Biochemistry and Molecular Biology
Record Identifier: 9984024543502771

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