We measured the temperature-dependent electronic circular dichroism (ECD) spectra of AX, XA, and XG dipeptides in D2O. The spectra of all XA and AX peptides indicate a substantial population of the polyproline II (PPII) conformation, while the ECD spectra of LG, KG, PG, and AG were found to be quantitatively different from the alanine-based dipeptides. Additional UV absorption data indicate that the ECD spectra of the XG peptides stem from electronic coupling between the peptide and the C-terminal group, and that spectral differences reflect different orientations of the latter. We also measured the H-1 NMR spectra of the investigated dipeptides to determine the (3)J(HaNH) coupling constants for the C-terminal residue. The observed temperature dependence of the ECD spectra and the respective room-temperature (3)J(HaNH) coupling constants were analyzed by it two-state model encompassing PPII and a beta-like coil formation. The PPII propensity of alanine in the XA series is only slightly modulated by the N-terminal side chain, and is larger than 50%. As compared to AA, XA peptides containing L, P, S, K V, E, T. and I all cause a relative stabilization of the extended beta-strand conformation. The PPII fractions of XA peptides varied between 0.64 for AA and 0.58 for DA, whereas the PPII fractions of AX peptides were much lower. From the investigated AX peptides. only AL and AQ showed the expected PPII propensity. We found that AT, AI. and AV clearly prefer an extended beta-strand conformation. A quantitative comparison of AA, AAA, and AAAA revealed a hierarchy AAAA > AAA approximate to AA for the PPII population, in agreement with predictions from MD calculations and results from Raman optical activity studies.