Loss of Cellular Sialidases Does Not Affect the Sialylation Status of the Prion Protein but Increases the Amounts of Its Proteolytic Fragment C1

The central molecular event underlying prion diseases involves conformational change of the cellular form of the prion protein (PrP C ), which is a sialoglycoprotein, into the disease-associated, transmissible form denoted PrP Sc . Recent studies revealed a correlation between the sialylation status of PrP Sc and incubation time to disease and introduced a new hypothesis that progression of prion diseases could be controlled or reversed by altering the sialylation level of PrP C . Of the four known mammalian sialidases, the enzymes that cleave off sialic acid residues, only NEU1, NEU3 and NEU4 are expressed in the brain. To test whether cellular sialidases control the steady-state sialylation level of PrP C and to identify the putative sialidase responsible for desialylating PrP C , we analyzed brain-derived PrP C from knockout mice deficient in Neu1 , Neu3 , Neu4 , or from Neu3/Neu4 double knockouts. Surprisingly, no differences in the sialylation of PrP C or its proteolytic product C1 were noticed in any of the knockout mice tested as compared to the age-matched controls. However, significantly higher amounts of the C1 fragment relative to full-length PrP C were detected in the brains of Neu1 knockout mice as compared to WT mice or to the other knockout mice. Additional experiments revealed that in neuroblastoma cell line the sialylation pattern of C1 could be changed by an inhibitor of sialylatransferases. In summary, this study suggests that targeting cellular sialidases is apparently not the correct strategy for altering the sialylation levels of PrP C , whereas modulating the activity of sialylatransferases might offer a more promising approach. Our findings also suggest that catabolism of PrP C involves its α- cleavage followed by desialylation of the resulting C1 fragments by NEU1 and consequent fast degradation of the desialylated products.