Double-stranded DNA must disentangle itself into single strands during replication or repair to allow functional molecules to bind and perform their various operations. Cellular proteins specifically bind to single-stranded DNA to prevent their premature recombination.
Unfortunately, detailed studies of these DNA–protein interactions have been hindered by the need for expensive instrumentation and time-consuming labelling techniques. Yen Nee Tan at the A*STAR Institute of Materials Research and Engineering and co-workers1 have now developed a convenient method to characterize the interactions between single-stranded DNA and their binding proteins.
The researchers used the optical properties of gold nanoparticles to probe the mechanism of protein–DNA binding. When the nanoparticles were well dispersed in solution, they yielded a bright red color, but when aggregated, the solution changed to blue. Tan and co-workers discovered that when single-stranded DNA and its binding protein were both present in the solution, coupled with a salt that stimulates nanoparticle aggregation, the DNA remained red in color, indicating that the DNA–protein complexes had bound with the nanoparticles through electrosteric stabilization forces. In contrast, when the protein or single-stranded DNA was introduced alone in the salt solution, there was a greater shift to the blue-grey color, indicating nanoparticle aggregation (see image).
“The greatest challenge in this work was to determine the optimum conditions for single-stranded DNA to bind with its binding protein to form complexes that confer the highest stability to gold nanoparticles from salt-induced aggregation,” says Tan.
The researchers attribute binding of the nanoparticles and the DNA–protein complexes to the presence of sulphur-containing groups in the protein, which are known to create strong bonds with gold. The protein molecules alone are smaller in molecular size than the protein–DNA complexes, leading to a less effective steric stabilization of the nanoparticles.
Tan and co-workers showed that there was a minimum length of DNA sequence under which the binding protein–DNA adhesion mechanism could operate. They found that the binding protein had a preference for binding to specific chemical units (bases) which make up DNA, and were able to spot DNA sequence variations, called single nucleotide polymorphisms (SNPs), even at the extreme ends of the molecule which are difficult to identify. Double-stranded DNA with SNPs cannot bind together so closely. The binding protein can thus attach to the dissociated single-stranded DNA to form protein–DNA complexes, offering sites to which gold nanoparticles can adhere.
“We plan to further develop this assay into a hassle-free genotyping assay to detect SNPs in real biological samples containing long genomic DNA,” says Tan.
The A*STAR-affiliated researchers contributing to this research are from the Institute of Materials Research and Engineering
Further reports about: > DNA > DNA sequence > DNA–protein > Ferchau Engineering > Nanotechnology > Probing > SNP > double-stranded DNA > gold nanoparticle > materials research > protein complexes > protein molecule > protein–DNA complexes > single nucleotide polymorphism > single nucleotide polymorphisms > single-stranded DNA
Ion treatments for cardiac arrhythmia — Non-invasive alternative to catheter-based surgery
20.01.2017 | GSI Helmholtzzentrum für Schwerionenforschung GmbH
Seeking structure with metagenome sequences
20.01.2017 | DOE/Joint Genome Institute
An important step towards a completely new experimental access to quantum physics has been made at University of Konstanz. The team of scientists headed by...
Yersiniae cause severe intestinal infections. Studies using Yersinia pseudotuberculosis as a model organism aim to elucidate the infection mechanisms of these...
Researchers from the University of Hamburg in Germany, in collaboration with colleagues from the University of Aarhus in Denmark, have synthesized a new superconducting material by growing a few layers of an antiferromagnetic transition-metal chalcogenide on a bismuth-based topological insulator, both being non-superconducting materials.
While superconductivity and magnetism are generally believed to be mutually exclusive, surprisingly, in this new material, superconducting correlations...
Laser-driving of semimetals allows creating novel quasiparticle states within condensed matter systems and switching between different states on ultrafast time scales
Studying properties of fundamental particles in condensed matter systems is a promising approach to quantum field theory. Quasiparticles offer the opportunity...
Among the general public, solar thermal energy is currently associated with dark blue, rectangular collectors on building roofs. Technologies are needed for aesthetically high quality architecture which offer the architect more room for manoeuvre when it comes to low- and plus-energy buildings. With the “ArKol” project, researchers at Fraunhofer ISE together with partners are currently developing two façade collectors for solar thermal energy generation, which permit a high degree of design flexibility: a strip collector for opaque façade sections and a solar thermal blind for transparent sections. The current state of the two developments will be presented at the BAU 2017 trade fair.
As part of the “ArKol – development of architecturally highly integrated façade collectors with heat pipes” project, Fraunhofer ISE together with its partners...
19.01.2017 | Event News
10.01.2017 | Event News
09.01.2017 | Event News
20.01.2017 | Awards Funding
20.01.2017 | Materials Sciences
20.01.2017 | Life Sciences