6. Isothermal Titration Calorimetry (ITC)

The ITC studies were conducted with a VP-ITC calorimeter (MicroCal, Northampton, MA). Prior to conducting the ITC studies, the dsDNA oligonucleotide and the JCV T-ag OBD proteins (both wt and the F258L mutant) were buffer-exchanged, using PD-10 columns (GE Healthcare), into 10 mM Sodium Phosphate, pH 7.0, 50 mM NaCl and 5 mM DTT. Protein and DNA concentrations were determined spectrophotometrically, using extinction coefficients calculated with the ProtParam web server and the Integrated DNA Technologies (IDT) website, respectively. The data were analyzed using the Origin software provided by the manufacturer.