Protein-immobilized electrode for rapid and convenient sensing of thyroid hormone receptor-ligand interaction.
The recombinant human thyroid hormone receptor (TR) was expressed as an in-frame fusion with ten consecutive histidine residues using a bacterial system; then the receptor was immobilized on an Au-electrode with Ni(II)-mediated chemisorption using the histidine tag and thiol-modified nitrilotriacetic acid. The receptor-modified electrode could rapidly detect ligand binding to hTR without any separation procedures, and showed a good response in a concentration-dependent manner. The sensitivity of this biosensor based on ligand-receptor interactions was comparable to those of conventional competitive ligand binding assays using radio-labeled ligands. Our results strongly suggest that our new biosensor can be applied to the identification of new ligands for hTR.
Murata
Masaharu
M
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka 812-8581, Japan.
Yano
Kentaro
K
Kuroki
Shinichiro
S
Suzutani
Tatsuo
T
Katayama
Yoshiki
Y
D015374:Biosensing Techniques / Q000379:methods*; D004566:Electrodes; D006046:Gold; D006639:Histidine; D006801:Humans; D008024:Ligands; D011485:Protein Binding; D011506:Proteins; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D013963:Thyroid Hormones / Q000032:analysis*
D008024:Ligands; D011506:Proteins; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D013963:Thyroid Hormones; D006639:Histidine; D006046:Gold