Red, Orange, Green: Light- and Temperature-Dependent Color Tuning in a Cyanobacteriochrome

Biochemistry. 2020 Feb 4;59(4):509-519. doi: 10.1021/acs.biochem.9b00931. Epub 2019 Dec 26.

Abstract

Cyanobacteriochromes (CBCRs) are photoreceptor proteins that photoconvert between two parent states and thereby regulate various biological processes. An intriguing property is their variable ultraviolet-visible (UV-vis) absorption that covers the entire spectral range from the far-red to the near-UV region and thus makes CBCRs promising candidates for optogenetic applications. Here, we have studied Slr1393, a CBCR that photoswitches between red- and green-absorbing states (Pr and Pg, respectively). Using UV-vis absorption, fluorescence, and resonance Raman (RR) spectroscopy, a further orange-absorbing state O600 that is in thermal equilibrium with Pr was identified. The different absorption properties of the three states were attributed to the different lengths of the conjugated π-electron system of the phycocyanobilin chromophore. In agreement with available crystal structures and supported by quantum mechanics/molecular mechanics (QM/MM) calculations, the most extended conjugation holds for Pr whereas it is substantially reduced in Pg. Here, the two outer pyrrole rings D and A are twisted out of the plane defined by inner pyrrole rings B and C. For the O600 state, the comparison of the experimental RR spectra with QM/MM-calculated spectra indicates a partially distorted ZZZssa geometry in which ring A is twisted while ring D and the adjacent methine bridge display essentially the same geometry as Pr. The quantitative analysis of temperature-dependent spectra yields an enthalpy barrier of ∼30 kJ/mol for the transition from Pr to O600. This reaction is associated with the movement of a conserved tryptophan residue from the chromophore binding pocket to a solvent-exposed position.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Color
  • Cyanobacteria / chemistry
  • Cyanobacteria / metabolism
  • Light
  • Molecular Dynamics Simulation
  • Photoreceptors, Microbial / chemistry*
  • Photoreceptors, Microbial / metabolism
  • Phycobilins / chemistry*
  • Phycobilins / metabolism
  • Phycocyanin / chemistry*
  • Phycocyanin / metabolism
  • Phycocyanin / ultrastructure
  • Phytochrome / chemistry
  • Pigments, Biological / chemistry
  • Synechocystis / chemistry*
  • Synechocystis / metabolism
  • Temperature

Substances

  • Bacterial Proteins
  • Photoreceptors, Microbial
  • Phycobilins
  • Pigments, Biological
  • Phycocyanin
  • Phytochrome
  • phycocyanobilin