Multicomponent Bioluminescence Imaging with a π-Extended Luciferin

Zi Yao, Brendan S. Zhang, Rachel C. Steinhardt, Jeremy H. Mills, Jennifer A. Prescher

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Bioluminescence imaging with luciferase-luciferin pairs is commonly used for monitoring biological processes in cells and whole organisms. Traditional bioluminescent probes are limited in scope, though, as they cannot be easily distinguished in biological environments, precluding efforts to visualize multicellular processes. Additionally, many luciferase-luciferin pairs emit light that is poorly tissue penetrant, hindering efforts to visualize targets in deep tissues. To address these issues, we synthesized a set of π-extended luciferins that were predicted to be red-shifted luminophores. The scaffolds were designed to be rotationally labile such that they produced light only when paired with luciferases capable of enforcing planarity. A luciferin comprising an intramolecular "lock"was identified as a viable light-emitting probe. Native luciferases were unable to efficiently process the analog, but a complementary luciferase was identified via Rosetta-guided enzyme design. The unique enzyme-substrate pair is red-shifted compared to well-known bioluminescent tools. The probe set is also orthogonal to other luciferase-luciferin probes and can be used for multicomponent imaging. Four substrate-resolved luciferases were imaged in a single session. Collectively, this work provides the first example of Rosetta-guided design in engineering bioluminescent tools and expands the scope of orthogonal imaging probes.

Original languageEnglish (US)
Pages (from-to)14080-14089
Number of pages10
JournalJournal of the American Chemical Society
Volume142
Issue number33
DOIs
StatePublished - Aug 19 2020

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Fingerprint

Dive into the research topics of 'Multicomponent Bioluminescence Imaging with a π-Extended Luciferin'. Together they form a unique fingerprint.

Cite this