Supramolecular Assembly of an Evolved Miniprotein Host and Fluorogenic Guest Pair

Bi Xu, Xinqi Zhou, Clifford I Stains

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Small-molecule-induced assembly of defined protein structures could have broad implications for the fabrication of new materials as well as biological signaling pathways. However, the design of new host-guest pairs capable of small-molecule-induced assembly in a biologically relevant context remains a significant challenge. Herein, we report a series of miniprotein hosts, evolved from the tenth type III domain of fibronectin (Fn3), that display remarkable binding affinity toward a red-shifted environment-sensitive merocyanine derivative, termed sI-Pht. Importantly, the consensus binder isolated from directed evolution experiments (6.2.18) forms a higher order assembly in response to addition of sI-Pht, as assessed by analytical ultracentrifugation. sI-Pht-induced assembly of 6.2.18 results in a 570-fold increase in fluorescence compared to free dye. This property enables the direct visualization of host-guest assemblies by fluorescence microscopy. As a demonstration, we show that supramolecular assembly of the 6.2.18-sI-Pht system can be visualized on the surface of living yeast cells. This new host-guest pair provides a tool for the potential development of new materials as well as pathway engineering. In a broader context, this work details a new design paradigm for the discovery of host-guest systems that function in the context of living cells.

Original languageEnglish (US)
Pages (from-to)14252-14255
Number of pages4
JournalJournal of the American Chemical Society
Volume137
Issue number45
DOIs
StatePublished - Nov 2 2015

Fingerprint

Ultracentrifugation
Fluorescence Microscopy
Coloring Agents
Yeasts
Fluorescence
Cells
Proteins
Molecules
Fluorescence microscopy
Fibronectins
Yeast
Biological materials
Binders
Demonstrations
Visualization
Dyes
Derivatives
Fabrication
Fibronectin Type III Domain
merocyanine

ASJC Scopus subject areas

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

Cite this

Supramolecular Assembly of an Evolved Miniprotein Host and Fluorogenic Guest Pair. / Xu, Bi; Zhou, Xinqi; Stains, Clifford I.

In: Journal of the American Chemical Society, Vol. 137, No. 45, 02.11.2015, p. 14252-14255.

Research output: Contribution to journalArticle

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