Kinetic and stoichiometric analysis for the binding of Escherichia coli ribonuclease HI to RNA-DNA hybrids using surface plasmon resonance.

@article{Haruki1997KineticAS,
  title={Kinetic and stoichiometric analysis for the binding of Escherichia coli ribonuclease HI to RNA-DNA hybrids using surface plasmon resonance.},
  author={Mitsuru Haruki and Eishi Noguchi and Shigenori Kanaya and Robert J Crouch},
  journal={The Journal of biological chemistry},
  year={1997},
  volume={272 35},
  pages={22015-22}
}
To understand how ribonucleases H recognize RNA-DNA hybrid substrates, we analyzed kinetic parameters of binding of Escherichia coli RNase HI to RNA-DNA hybrids ranging in length from 18 to 36 base pairs (bp) using surface plasmon resonance (BIAcoreTM). The kon and koff values for the binding of the enzyme to the 36-bp substrate were 1.5 x 10(6) M-1 s-1 and 3.2 x 10(-2) s-1, respectively. Similar values were obtained with the shorter substrates. Using uncleavable 2'-O-methylated RNA-DNA… CONTINUE READING