Generation and characterisation of C-terminally stabilised PYY molecules with potential in vivo NPYR2 activity

Abstract

BackgroundActivation of neuropeptide Y2 receptors (NPYR2) by the N-terminally truncated, dipeptidyl peptidase-4 (DPP-4) generated, Peptide YY (PYY) metabolite, namely PYY(3–36), results in satiating actions. However, PYY(3–36) is also subject to C-terminal enzymatic cleavage, which annuls anorectic effects. MethodsSubstitution of l-Arg35 with d-Arg35 in the DPP-4 stable sea lamprey PYY(1–36) peptide imparts full C-terminal stability. In the current study, we have taken this molecule and introduced DPP-4 susceptibility by Iso3 substitution. ResultsAs expected, [Iso3]sea lamprey PYY(1–36) and [Iso3](d-Arg35)sea lamprey PYY(1–36) were N-terminally degraded to respective PYY(3–36) metabolites in plasma. Only [Iso3](d-Arg35)sea lamprey PYY(1–36) was C-terminally stable. Both peptides possessed similar insulinostatic and anti-apoptotic biological actions to native PYY(1–36) in beta-cells. Unlike native PYY(1–36) and [Iso3](d-Arg35)sea lamprey PYY(1–36), [Iso3]sea lamprey PYY(1–36) displayed some proliferative actions in Npyr1 knockout beta-cells. In addition, [Iso3]sea lamprey PYY(1–36) induced more rapid NPYR2-dependent appetite suppressive effects in mice than its C-terminally stable counterpart. Twice daily administration of either peptide to high fat fed (HFF) mice resulted in significant body weight reduction and improvements in circulating triglyceride levels. [Iso3]sea lamprey PYY(1–36) treatment also prevented elevations in glucagon. Both peptides, and especially [Iso3]sea lamprey PYY(1–36), improved glucose tolerance. The treatment interventions also partially reversed the deleterious effects of sustained high fat feeding on pancreatic islet morphology. ConclusionThe present study confirms that sustained NPYR2 receptor activation by [Iso3](d-Arg35)sea lamprey induced significant weight lowering actions. However, identifiable benefits of this peptide over [Iso3]sea lamprey PYY(1–36), which was not protected against C-terminal degradation, were not pronounced.