Thermodynamic characterization of specific interactions between the human Lon protease and G-quartet DNA
Resource
Nucleic Acids Research 36 (4): 1273-1287
Journal
Nucleic Acids Research
Pages
1273-1287
Date Issued
2008
Date
2008
Author(s)
Chen, Si-Han
Suzuki, Carolyn K.
Wu, Shih-Hsiung
Abstract
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.
SDGs
File(s)![Thumbnail Image]()
Loading...
Name
77.pdf
Size
760.44 KB
Format
Adobe PDF
Checksum
(MD5):f78804d5517fa35ed2127757066135e3