Nucleic Acids Research Advance Access originally published online on January 3, 2008
Nucleic Acids Research 2008 36(4):1273-1287; doi:10.1093/nar/gkm1140
Nucleic Acids Research, 2008, Vol. 36, No. 4 1273-1287
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Structural Biology |
Thermodynamic characterization of specific interactions between the human Lon protease and G-quartet DNA
1Institute of Biological Chemistry, Academia Sinica, Taipei 115, 2Institute of Biochemical Science, National Taiwan University, Taipei 106, Taiwan and 3Department of Biochemistry and Molecular Biology, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103, USA
*To whom correspondence should be addressed. Tel: +886 2 27855696 7101; Fax: +886 2 27889759; Email: shwu{at}gate.sinica.edu.tw
Received September 5, 2007. Revised November 24, 2007. Accepted December 7, 2007.
Lon is an ATP-powered protease that binds DNA. However, the function of DNA binding by Lon remains elusive. Studies suggest that human Lon (hLon) binds preferentially to a G-rich single-stranded DNA (ssDNA) sequence overlapping the light strand promoter of mitochondrial DNA. This sequence is contained within a 24-base oligonucleotide referred to as LSPas. Here, we use biochemical and biophysical approaches to elucidate the structural properties of ssDNAs bound by hLon, as well as the thermodynamics of DNA binding by hLon. Electrophoretic mobility shift assay and circular dichroism show that ssDNAs with a propensity for forming parallel G-quartets are specifically bound by hLon. Isothermal titration calorimetry demonstrates that hLon binding to LSPas is primarily driven by enthalpy change associated with a significant reduction in heat capacity. Differential scanning calorimetry pinpoints an excess heat capacity upon hLon binding to LSPas. By contrast, hLon binding to an 8-base G-rich core sequence is entropically driven with a relatively negligible change in heat capacity. A considerable enhancement of thermal stability accompanies hLon binding to LSPas as compared to the G-rich core. Taken together, these data support the notion that hLon binds G-quartets through rigid-body binding and that binding to LSPas is coupled with structural adaptation.