Site-specific analysis of N-glycans from different sheep prion strains

Abstract

Prion diseases are a group of neurodegenerative diseases affecting a wide range of mammalian species, including humans. During the course of the disease, the abnormally folded scrapie prion protein (PrP^Sc) accumulates in the central nervous system where it causes neurodegeneration. In prion disorders, the diverse spectrum of illnesses exists because of the presence of different isoforms of PrP^Sc where they occupy distinct conformational states called strains. Strains are biochemically distinguished by a characteristic three-band immunoblot pattern, defined by differences in the occupancy of two glycosylation sites on the prion protein (PrP). Characterization of the exact N-glycan structures attached on either PrP^C or PrP^Sc is lacking. Here we report the characterization and comparison of N-glycans from two different sheep prion strains. PrP^Sc from both strains was isolated from brain tissue and enzymatically digested with trypsin. By using liquid chromatography coupled to electrospray mass spectrometry, a site-specific analysis was performed. A total of 100 structures were detected on both glycosylation sites. The N-glycan profile was shown to be similar to the one on mouse PrP, however, with additional 40 structures reported. The results presented here show no major differences in glycan composition, suggesting that glycans may not be responsible for the differences in the two analyzed prion strains. To date, prion diseases remain a controversy amongst scientists. Although we know now it is the abnormal form of the prion protein (PrP^Sc) that causes the disease, many questions are still left unanswered. To understand the cellular mechanism of these diseases, we should first and foremost try to fully understand the prion protein itself. Even though many findings have been made regarding the structure of the protein, a large part of it is still unknown. Since the prion protein is actually a glycoprotein, to resolve its structure we need to put our focus not only on the protein part of the glycoprotein but also on the glycan structures as well. Here we compared two different sheep prion strains and although no major differences have been found between the glycan structures, this analysis may help the understanding of the role glycans have in prion diseases.