Nano-mechanical mapping of the interactions between surface-bound RC-LH1-PufX core complexes and cytochrome c(2) attached to an AFM probe

Electron transfer pathways in photosynthesis involve interactions between membrane-bound complexes such as reaction centres with an extrinsic partner. In this study, the biological specificity of electron transfer between the reaction centre-light-harvesting 1-PufX complex and its extrinsic electron donor, cytochrome c(2), formed the basis for mapping the location of surface-attached RC-LH1-PufX complexes using atomic force microscopy (AFM). This nano-mechanical mapping method used an AFM probe functionalised with cyt c(2) molecules to quantify the interaction forces involved, at the single-molecule level under native conditions. With surface-bound RC-His(12)-LH1-PufX complexes in the photo-oxidised state, the mean interaction force with cyt c(2) is approximately 480 pN with an interaction frequency of around 66 %. The latter value lowered 5.5-fold when chemically reduced RC-His(12)-LH1-PufX complexes are imaged in the dark to abolish electron transfer from cyt c(2) to the RC. The correspondence between topographic and adhesion images recorded over the same area of the sample shows that affinity-based AFM methods are a useful tool when topology alone is insufficient for spatially locating proteins at the surface of photosynthetic membranes.