Detection of protein-DNA interaction and regulation using gold nanoparticles.
The interaction between protein and DNA is usually regulated by a third species, an effector, which can be either a protein or a small molecule. Convenient methods capable of detecting protein-DNA interaction and its regulation are highly desirable research tools. In the current study, we developed a method to directly "visualize" the interaction between a protein-DNA pair and its effector through the coupling with gold nanoparticles (AuNPs). As a proof-of-concept experiment, we constructed a model system based on the interaction between the lac repressor (protein) and operator (DNA) and its interplay with the lac operon inducer isopropyl beta-D-1-thiogalactopyranoside (IPTG, which inhibits the interaction between the lac repressor and operator). We coated AuNPs with the lac operator sequences and mixed them with the lac repressor. Because the lac repressor homotetramer contains two DNA binding modules, it bridged the particles and caused them to aggregate. We demonstrated that the assembly of DNA-modified AuNPs correlated with the presence of the corresponding protein and effector in a concentration-dependent manner. This AuNP-based platform has the potential to be generalized in the creation of reporter and detection systems for other interacting protein-DNA pairs and their effectors.
Fang
Jun
J
Department of Chemistry, University of Kentucky, Lexington, KY 40506, USA.
Yu
Linliang
L
Gao
Pei
P
Cai
Yuguang
Y
Wei
Yinan
Y
D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D056968:Lac Repressors / Q000235:genetics / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry*; D018625:Microscopy, Atomic Force / Q000379:methods*; D011485:Protein Binding
D004268:DNA-Binding Proteins; D056968:Lac Repressors; D007544:Isopropyl Thiogalactoside; D006046:Gold; D004247:DNA