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α2,3 linkage of sialic acid to a GPI anchor and an unpredicted GPI attachment site in human prion protein.

ABSTRACT: Prion diseases are transmissible, lethal neurodegenerative disorders caused by accumulation of the aggregated scrapie form of the prion protein (PrP^Sc) after conversion of the cellular prion protein (PrP^C). The glycosylphosphatidylinositol (GPI) anchor of PrP^C is involved in prion disease pathogenesis, and especially sialic acid in a GPI side chain reportedly affects PrP^C conversion. Thus, it is important to define the location and structure of the GPI anchor in human PrP^C Moreover, the sialic acid linkage type in the GPI side chain has not been determined for any GPI-anchored protein. Here we report GPI glycan structures of human PrP^C isolated from human brains and from brains of a knock-in mouse model in which the mouse prion protein (Prnp) gene was replaced with the human PRNP gene. LC-electrospray ionization-MS analysis of human PrP^C from both biological sources indicated that Gly²²⁹ is the ω site in PrP^C to which GPI is attached. Gly²²⁹ in human PrP^C does not correspond to Ser²³¹, the previously reported ω site of Syrian hamster PrP^C We found that ∼41% and 28% of GPI anchors in human PrP^Cs from human and knock-in mouse brains, respectively, have N-acetylneuraminic acid in the side chain. Using a sialic acid linkage-specific alkylamidation method to discriminate α2,3 linkage from α2,6 linkage, we found that N-acetylneuraminic acid in PrP^C's GPI side chain is linked to galactose through an α2,3 linkage. In summary, we report the GPI glycan structure of human PrP^C, including the ω-site amino acid for GPI attachment and the sialic acid linkage type.

SUBMITTER: Kobayashi A

PROVIDER: S-EPMC7261787 | biostudies-literature |

REPOSITORIES: biostudies-literature

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