Although the mechanism of the conformational conversion from the cellular (PrP(C)) to the scrapie (PrP(Sc)) form of animal prion proteins has yet to be elucidated, evidence is accumulating that may provide insight into the conversion process at atomic resolution. Here we show critical aspects of the slow fluctuation dynamics of the recombinant hamster prion protein, rPrP(90-231), based on NMR relaxation analysis using Carr-Purcell-Meiboom-Gill (CPMG) experiments, and compare them in detail with results from high-pressure NMR. Residues exhibiting slow fluctuations on the time scale of microseconds to milliseconds are mainly localized on helices B and C (172-193 and 200-227), which include locally disordered regions in an intermediate conformer, PrP*, identified previously by high-pressure NMR [Kuwata, K., et al., (2002) Biochemistry 41, 12277-12283]. Moreover, chemical shift differences between two putative exchanging conformers obtained by the CPMG relaxation analysis and the linear component of the pressure-induced chemical shift changes are reasonably correlated at individual residue sites. These observations suggest that both the CMPG relaxation and the pressure shifts reflect slow conformational fluctuations and that these slow motions in PrP(C) are related to the trajectories leading to the transition to PrP*.