Novel biosensor using split-luciferase for detecting vitamin D receptor ligands based on the interaction between vitamin D receptor and coactivator.

Abstract

Vitamin D receptor (VDR) ligands, such as 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] and its analogs, have been investigated for their potential clinical use in the treatment of various diseases such as type I rickets, osteoporosis, psoriasis, leukemia, and cancer. Previously, we reported a split-luciferase-based biosensor that can detect VDR ligands and assess their affinity for the ligand binding domain (LBD) of the VDR in a short time. However, a further increase in its sensitivity was required to detect plasma levels of 1α,25(OH)2D3 and its analogs. In this study, a novel type of biosensor called LXXLL + LBD was successfully developed. Here, the split luciferase forms a functional complex based on the intermolecular interaction between the LXXLL motif and the ligand-bound form of the LBD. This biosensor has an approximately 10-fold increase in the light intensity compared to the previous versions. Additionally, the binding affinity of the vitamin D analogs for the wild-type and the rickets-associated mutant R274L of VDR was evaluated.

• Authors: Hiroki Mano, Masashi Takano, Shinichi Ikushiro, Atsushi Kittaka, Toshiyuki Sakaki
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 505 Issue 2 Pg. 460-465 (10 28 2018) ISSN: 1090-2104 [Electronic] United States
• PMID: 30268505 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2018 Elsevier Inc. All rights reserved.

Chemical References

• Ligands
• Receptors, Calcitriol
• dihydroxy-vitamin D3
• Vitamin D
• Luciferases

Topics

• Amino Acid Motifs
• Biosensing Techniques (instrumentation, methods)
• Humans
• Ligands
• Luciferases
• Protein Binding
• Protein Domains
• Receptors, Calcitriol (genetics, metabolism)
• Rickets (diagnosis)
• Vitamin D (analogs & derivatives, analysis)