Molecular imaging of retinoic acids in live cells using single-chain bioluminescence probes

Sung Bae Kim, Rika Fujii, Ryo Nishihara, Rajendran J.C. Bose, Daniel Citterio, Koji Suzuki, Tarik F. Massoud, Ramasamy Paulmurugan

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Retinoic acid (RA) is a key metabolite necessary for embryonic development and differentiation in vertebrates. We demonstrate the utility of genetically encoded, ligand-activatable single-chain bioluminescence probes for detecting RAs from different biological sources. We examined 13 different molecular designs to identify an efficient single-chain probe that can quantify RA with significant sensitivity. The optimal probe consisted of four components: the N- and C-terminal fragments of artificial luciferase variant-16 (ALuc16), the ligand binding domain of retinoic acid receptor α (RARα LBD), and an LXXLL interaction motif. This probe showed a 5.2-fold greater bioluminescence intensity in response to RA when compared to the vehicle control in live mammalian cells. The probe was highly selective to all-trans-RA (at-RA), and highly sensitive in determining at-RA levels in cells derived from tumor xenografts created using MDA-MB-231 cells engineered to stably express the probe. We also detected RA levels in serum and cerebrospinal fluid. Using this probe, the detection limit for at-RA was 10-9.5 M, with a linear range of two orders. We present a highly useful technique to quantitatively image endogenous at-RA levels in live mammalian cells expressing novel single-chain bioluminescence probes.

Original languageEnglish
Pages (from-to)473-481
Number of pages9
JournalACS Combinatorial Science
Volume21
Issue number6
DOIs
Publication statusPublished - 2019 Jun 10

Keywords

  • Artificial luciferase
  • Bioluminescence
  • Protein-protein interactions
  • Retinoic acid receptor
  • Single-chain probe
  • retinoic acids

ASJC Scopus subject areas

  • Chemistry(all)

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