Evaluation of the Influence of Biosurface Design on the Interaction between the Regulatory Peptide RS1-reg and ODC1 Reveals a Membrane-Dependent Affinity Increase

The regulatory solute carrier protein, family 1, member 1 (RS1) modulates via its N-terminal domain RS1-reg the activity of Na+-d-glucose cotransporter 1 (SGLT1) and thereby the glucose uptake in the small intestine by blocking the release of SGLT1-containing vesicles at the trans-Golgi network (TGN). The antidiabetic activity of RS1 is mediated by ornithindecarboxylase 1 (ODC1), catalyzing the conversion of ornithine to putrescine. Putrescine can bind to a buddying protein complex for SGLT1-containing vesicles at the membrane of the TGN, triggering vesicle release. In this report, a first in-depth analysis of the important binding process between ODC1 and RS1-reg for regulating glucose uptake in the human organism is described by comparing results from the surface-based methods, "surface plasmon resonance" (SPR) and "surface acoustic wave" (SAW) with findings by isothermal titration calorimetry (ITC). In cases of SAW and SPR, three different assay surface setups are compared, resulting in small but significant differences in K-D values for different surfaces. Noteworthy, an affinity increase by the factor of about 100 for the interaction is detected and herewith described for the first time in the presence of biological membranes that may be relevant in vivo for the biological function of RS1 and future bespoken antidiabetic drug development.