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Photoswitchable affinity reagents: Computational design and efficient red-light switching.

Nobuo Yasuike1,2, Kristin M Blacklock3, Huixin Lu1

  • 1Department of Chemistry, University of Toronto, 80 St. George St., Toronto, M5S 3H6, Canada.

Chemphotochem
|August 29, 2020
PubMed
Summary

Researchers explored photo-controlled affinity reagents for spatiotemporal bioactivity control. They computationally screened and experimentally tested photoswitchable cross-linkers on Fynomer scaffolds, observing limited photo-switchability.

Keywords:
azobenzeneazoniumfynomeroptogeneticphotopharmacology

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Biology

Background:

  • Photo-controlled affinity reagents offer spatiotemporal regulation of biological activity.
  • Developing modular systems for precise control of bioactivity is crucial in molecular biology.

Purpose of the Study:

  • To computationally screen and experimentally validate sites for attaching photoswitchable cross-linkers to the Fynomer affinity reagent.
  • To assess the photo-switchability and binding affinity of modified Fynomers using UV-based and red-light switchable cross-linkers.

Main Methods:

  • Utilized Rosetta-based computational methods to identify optimal attachment sites on the Fynomer structure.
  • Experimentally tested azobenzene-iodoacetamide (IAC) and piperazino-tetra-ortho-methoxy azobenzene (PIP) cross-linkers.
  • Evaluated Fynomer function and photo-switchability under UV and red light (633 nm).

Main Results:

  • Identified several attachment sites on the Fynomer compatible with cross-linker function.
  • Observed rapid (<10s) red light modulation of Fynomer function at specific sites.
  • Reported generally small photo-switchability (<2-fold) in Fynomer function across tested designs.

Conclusions:

  • Computational and experimental approaches identified viable sites for photoswitchable cross-linker attachment on Fynomers.
  • The degree of photo-switchability achieved was limited, potentially due to local flexibility.
  • Further optimization is needed to enhance the photo-switchability of these affinity reagents.