Photocycle characterization of a blue-orange cyanobacteriochrome from Synechococcus sp. PCC 7002

Abstract

Cyanobacteria employ photoreceptors called cyanobacteriochromes (CBCRs) to sense the colour and intensity of light. The information extracted from the solar spectrum is used for adaptive responses such as optimizing photosynthesis, phototaxis and cell aggregation. cGMP-phosphodiesterase/adenlylate cyclase/FhlA (GAF) domains are the principal light sensors in cyanobacteriochromes. They contain a conjugated bilin chromophore and boast an impressive spectral diversity. Characterizing the spectral characteristics of GAF domains in model strains, such as Synechococcus sp. PCC 7002, can open new avenues for optogenetics and biotechnology. Based on sequence analysis we predicted several different GAF domains in this strain. The SynPCC7002_a0852 gene encodes a single GAF domain with two cysteine residues: one in the conserved 3 helix and one in the conserved DXCF motif. Spectral analysis of recombinant SynPCC7002_A0852 with phycocyanobilin (PCB) showed that the protein cycles between two states, Po and Pb, which absorb orange and blue light, respectively. Measurements of kinetics identified Po as the dark state of the protein. Acid-denaturation analysis suggested that the 15E isomer of PCB is bound in the (dark) Po state, whereas 15Z is bound the (photoproduct) Pb state. Site-directed mutagenesis and iodoacetamide treatments showed that Cys73 in the DXCF motif is essential for the conversion from Po to Pb. Future experiments dark-purified protein/chromophore versions are required to establish the sequence of events in the photocycle. In summary, SynPCC7002_A0852 enables orange/blue colour perception in Synechococcus sp. PCC 7002 as other CBRCs of this protein family but might contain the energetically higher chromophore isoform in its dark state. Such photocycle has previously been found in bathy bacteriophytochromes but not in CBCRs.