Protein-induced changes in DNA structure and dynamics observed with noncovalent site-directed spin-labeling and PELDOR
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Site-directed spin labeling and pulsed electron–electron double resonance (PELDOR or DEER) have previously been applied successfully to study the structure and dynamics of nucleic acids. Spin labeling nucleic acids at specific sites requires the covalent attachment of spin labels, which involves rather complicated and laborious chemical synthesis. Here, we use a noncovalent label strategy that bypasses the covalent labeling chemistry and show that the binding specificity and efficiency are large enough to enable PELDOR or DEER measurements in DNA duplexes and a DNA duplex bound to the Lac repressor protein. In addition, the rigidity of the label not only allows resolution of the structure and dynamics of oligonucleotides but also the determination of label orientation and protein-induced conformational changes. The results prove that this labeling strategy in combination with PELDOR has a great potential for studying both structure and dynamics of oligonucleotides and their complexes with various ligands.
Reginsson , G W , Shelke , S , Rouillon , C , White , M F , Sigurdsson , S T & Schiemann , O 2013 , ' Protein-induced changes in DNA structure and dynamics observed with noncovalent site-directed spin-labeling and PELDOR ' Nucleic Acids Research , vol 41 , no. 1 , e11 . DOI: 10.1093/nar/gks817
Nucleic Acids Research
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