DNA binding kinetics of Oct-1 linker mutant proteins studied with the instrument for biomolecular interaction sensing (IBIS)

摘要

The binding constants of two proteins (a wild type and a mutant of Oct-i) that interact with a viral DNA are measured with a gel electrophoresis technique and with the Instrument for Biomolecular Interaction Sensing (IBIS). IBIS enables scientists to monitor biomolecular interactions in real-time and label-free for kinetic analysis of obtained interaction plots. The detection principle of IBIS is Surface Plasmon Resonance (SPR). IBIS detects changes in refractive index at the sensor surface. Thesechanges are directly related to the amount of sensor surface-bound biomolecules. The SPR-measurements with the protein Oct-i confirm biochemical band shifts measurements showing a difference in binding affinity between the wild-type and mutant Oct-iprotein (2.9 fold measured from IBIS; 2.5 fold from gel-electrophoreses experiments).