Publication date: 8th October 2020
It is challenging to study the nanoscale biophysics the proteins within intact thylakoid membranes because they are complicated, high-density, multi-layer systems. Here, we present “bio-hybrid membranes” which are generated by combining natural thylakoids and synthetic lipids and assembling them into a micro-patterned template of polymerized lipids. This platform has several useful properties including: a native-like mixture of membrane proteins, a facile self-assembly process, and a robust template which promotes the formation of supported lipid bilayers from such high-protein-density natural membranes. These hybrid membranes allowed us to assess how the energy transfer pathways and protein arrangement are correlated. Fluorescence Lifetime Imaging Microscopy revealed that the fluorescence lifetime of the light-harvesting (LH) proteins within hybrid membranes is significantly longer than native membranes and this, combined with Atomic Force Microscopy data, suggested that protein-protein interactions are reduced. The migration of LH proteins from natural membranes into the putative assembling hybrid membranes was observed in real time with fluorescence microscopy. Finally, hybrid membranes were compared to proteoliposomes as a basis to assess the potential electron activity of Photosystem II, however, these experiments revealed that common photochemical assays used in the photosynthesis community actually produce false-positive results for LH antenna complex proteins. A pre-print of this work is available here [1].
This collaboration between Leeds and Kobe was supported by an International Exchanges Cost Share award from The Royal Society UK (IEC\R3\183029) and Japan-UK Research Cooperative Program award from the Japan Society for the Promotion of Science (JPJSBP120195707). Other support included grants from Biotechnology and Biological Sciences Research Council (award numbers BB/M011151/1 and BB/R000174/1) the Engineering and Physical Sciences Research Council (award numbers 1807029, EP/ J017566/1, EP/P023266/1) and the Medical Research Council (MR/M009084/1).
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