From cytoplasm to lumen-mapping the free pools of protein subunits of three photosynthetic complexes using quantitative mass spectrometry.

The phycobilisome (PBS) captures light energy and transfers it to photosystem I (PSI) and photosystem II (PSII). Which and how many copies of protein subunits in PBSs, PSI, and PSII remain unbound in thylakoids are unknown. Here, quantitative mass spectrometry (QMS) was used to quantify substantial pools of free extrinsic subunits of PSII and PSI. Interestingly, the membrane intrinsic PsaL is 3-fold higher than PsaA/B. This scenario complements the static structures of these complexes as revealed by X-ray crystallography and cryo-EM. The ratios of ApcG and photoprotective OCP over PBS indicate a pool of extra ApcG. The 2.5 ratio of CpcG-PBS over CpcL-PBS improves our understanding of these light-harvesting complexes involved in energy capture and photoprotection in cyanobacteria. Impact statement Our study presents the first quantitative inquiry of the free pools of proteins associated with the three major photosynthetic complexes in Synechocystis 6803. This study increases our understanding of the unbound thylakoid proteome, guiding future research into the functions of these proteins, which will facilitate efforts to enhance photosynthetic efficiency.