Publication date: 8th October 2020
Photosystem I from the thermophilic cyanobacterium Thermosynechococcus elongatus is the largest membrane protein complex to have had its structure solved by X-ray diffraction. This trimeric complex has 51 transmembrane domains, 33 distinct subunits, over 330 non-covalently bound cofactors and a molecular weight of ~1.2 MDa. We have succeeded in isolating this complex without the use of detergents, using styrene maleic acid (SMA) alternating copolymer. The resulting SMA lipid particle (SMALP) containing trimeric PSI remains the largest protein complex to be isolated by this method. A host of biochemical, biophysical and functional assays have been applied to characterize this non-detergent form of PSI. Once formed, PSI-SMALP from T. elongatus have demonstrated a 3 nm red-shift in chlorophyll fluorescence, suggesting a more native orientation of the chlorophyll antennae within the PSI, as well as faster reduction kinetics following a photooxidation event. The latter observation may be related to an ultra-fast charge separation event that has been observed within PSI-SMALP by transient absorption femtosecond spectroscopy. This study shows a ~1,000-fold increase in the charge separation event of PSI-SMALP (~80 fs) over detergent solubilized complex (36 ps). Using the powerful tools of contrast variation and matching available only to SANS, we have performed a detailed dimensional analysis of the ~1.47 MDa PSI-SMALP, breaking it into its individual components of protein core, lipid annulus and copolymer or detergent micelle layers (for PSI-DDM). These trimeric PSI nanodiscs show ~50-fold increase in native lipids retained, with sulfoquinovosyldiacylglycerol being the most enriched lipid class in these PSI-SMALPs, suggesting the lipids of the thylakoid membrane are heterogeneously distributed, with alterations in the local profile within these highly uniform, PSI containing regions. These highly active and cost-effective PSI nanodiscs show great promise for scalability and integration into biohybrid solar devices for sustainable, non-resource limited electricity generation from sunlight.
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