Prion diseases are characterized biochemically by protein aggregation of infectious prion isoforms (PrP^Sc), which result from the conformational conversion of physiological prion proteins (PrP^C). PrP^C are variable post-translationally modified glycoproteins, which exist as full length and as aminoterminally truncated glycosylated proteins and which exhibit differential detergent solubility. This implicates the presence of heterogeneous phenotypes, which overlap as protein complexes at the same molecular masses. Although the biological function of PrP^C is still enigmatic, evidence reveals that PrP^C exhibits metal-binding properties, which result in structural changes and decreased solubility. In this study, we analyzed the yield of PrP^C metal binding affiliated with low solubility and changes in protein banding patterns. By implementing a high-speed centrifugation step, the interaction of zinc ions with PrP^C was shown to generate large quantities of proteins with low solubility, consisting mainly of full-length glycosylated PrP^C; whereas unglycosylated PrP^C remained in the supernatants as well as truncated glycosylated proteins which lack of octarepeat sequence necessary for metal binding. This effect was considerably lower when PrP^C interacted with copper ions; the presence of other metals tested exhibited no effect under these conditions. The binding of zinc and copper to PrP^C demonstrated differentially soluble protein yields within distinct PrP^C subtypes. PrP^C–Zn²⁺-interaction may provide a means to differentiate glycosylated and unglycosylated subtypes and offers detailed analysis of metal-bound and metal-free protein conversion assays.