Tethered Particle Motion Analysis of the DNA Binding Properties of Architectural Proteins

Architectural DNA binding proteins are key to the organization and compaction of genomic DNA inside cells. Tethered Particle Motion (TPM) permits analysis of DNA conformation and detection of changes in conformation induced by such proteins at the single molecule level in vitro. As many individual p...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1624; p. 127
Main Authors van der Valk, Ramon A, Laurens, Niels, Dame, Remus T
Format Journal Article
LanguageEnglish
Published United States 2017
Subjects
Biophysical Phenomena
DNA - chemistry
DNA - metabolism
DNA-Binding Proteins - metabolism
Motion
Nucleic Acid Conformation
Single Molecule Imaging - methods
Tethered particle motion
DNA stiffening
DNA conformation
DNA bending
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Summary:Architectural DNA binding proteins are key to the organization and compaction of genomic DNA inside cells. Tethered Particle Motion (TPM) permits analysis of DNA conformation and detection of changes in conformation induced by such proteins at the single molecule level in vitro. As many individual protein-DNA complexes can be investigated in parallel, these experiments have high throughput. TPM is therefore well suited for characterization of the effects of protein-DNA stoichiometry and changes in physicochemical conditions (pH, osmolarity, and temperature). Here, we describe in detail how to perform Tethered Particle Motion experiments on complexes between DNA and architectural proteins to determine their structural and biochemical characteristics.
ISSN:1940-6029
DOI:10.1007/978-1-4939-7098-8_11