AI Article Synopsis
- Cortisol is a key biomarker for stress, and researchers developed highly sensitive bioluminescent probes to detect salivary cortisol levels.
- The innovative design involved splitting Gaussia princeps luciferase and inserting a ligand binding domain for glucocorticoid receptor to create ON-OFF switches in a reporter-gene system.
- The final probes showed great improvements in detection limits and signal ratios, successfully tracking daily cortisol fluctuations and exhibiting correlations with traditional ELISA methods.
Article Abstract
Cortisol is a classical biomarker for the stress levels of human beings. We fabricated highly sensitive bioluminescent probes for salivary cortisol. The following strategies were contrived in the molecular design. Gaussia princeps luciferase (GLuc) was dissected into two fragments, between which an N-terminal-extended ligand binding domain of glucocorticoid receptor (GR HLBD), named Simgr4, was inserted. First, this unique single-chain probe was then situated downstream of a glucocorticoid response element (GRE) promoter in a reporter-gene system for constructing two ON-OFF switches for cortisol. Second, a circularly permutated (CP) variant of Simgr4 was formulated. The reporter-gene system exerted an improved signal-to-background (S/B) ratio of 8.5 to cortisol. Furthermore, a circularly permutated (CP) variant of Simgr4 exerted a 10× enhanced detection limit to cortisol and a long dynamic range from 10(-9) to 10(-6) M cortisol, covering all of the normal clinical ranges of serum, urine, and saliva. This optimized probe successfully determined daily fluctuations of salivary cortisol and the correlations with those by ELISA. This study is the first to investigate the contribution of the HLBD of a nuclear receptor and multiple ON-OFF switches for molecular probes and salivary cortisols.
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| http://dx.doi.org/10.1021/bc200220k | DOI Listing |
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