Figure 2.

PLA₂R interacts with A2t complex via S100A10. (a) Left, characterisation of Annexin2-S100A10 complex in solution using Multi Angle Laser Light Scattering (MALLS). The elution profile reveals 2 peaks, 110 kDa referred as A2t (Anxa2/S100A10 heterotetramer) and 47 kDa complex. Middle, 10 µg of purified complex ran on a non-reduced SDS-PAGE gel and stained with Instant Blue confirm the presence of both Anxa2 and S100A10 proteins in various oligomeric forms. Right, schematic representation of A2t complex made of a S100A10 dimer (10.2210/pdb1BT6/pdb) and two Anxa2 molecules (10.2210/pdb2hyw/pdb). (b) SPR data showing direct interaction with PLA₂R fragments and A2t complex. Purified PLA₂R NC8 and NC3 at various concentrations (0 to 300 nM) were injected over the surfaces of immobilised A2t. The maximal response (RU_max) were obtained from the sensograms (inset) and plotted against the increasing concentrations of injected PLA₂R. Each data point in both experiments is the mean of 3 repeats. The equilibrium dissociation constant, K _D, was calculated by fitting the data to the Hill equation using nonlinear regression. (c) Representative sensorgrams derived from injections of different concentrations of purified recombinant S100A10 protein over immobilised PLA₂R NC3. Results were obtained after reference subtraction.