Amino acid conditions near the GPI anchor attachment site of prion protein for the conversion and the GPI anchoring.

Amino acid conditions near the GPI anchor attachment site of prion protein for the conversion and the GPI anchoring.

Biochem Biophys Res Commun. 2009 Dec 26;

Authors: Hizume M, Kobayashi A, Mizusawa H, Kitamoto T

Prion protein (PrP) is a glycosylphosphatidylinositol (GPI) -anchored protein, and the C-terminal GPI anchor signal sequence (GPI-SS) of PrP is cleaved before GPI anchoring. However, mutations near the GPI anchor attachment site (the omega site) in the GPI-SS have been recognized in human genetic prion diseases. Moreover, the omega site of PrP has not been identified except hamster, though it is known that amino acid restrictions are very severe at the omega and omega + 2 sites in other GPI-anchored proteins. To investigate the effect of mutations near the omega site of PrP on the conversion and the GPI anchoring, and to discover the omega site of murine PrP, we systematically created mutant murine PrP with all possible single amino acid substitutions at every amino acid residue from codon 228 to 240. We transfected them into scrapie-infected mouse neuroblastoma cells and examined the conversion efficiencies and the GPI anchoring of each mutant PrP. Mutations near the omega site altered the conversion efficiencies and the GPI anchoring efficiencies. Especially, amino acid restrictions for the conversion and the GPI anchoring were severe at codons 230 and 232 in murine PrP, though they were less severe than in other GPI-anchored proteins. Only the mutant PrPs presented on a cell surface via a GPI anchor were conversion competent. The present study shows that mutations in the GPI-SS can affect the GPI anchoring and the conversion efficiency of PrP. We clarified for the first time the omega site of murine PrP and the amino acid conditions near the omega site for the conversion as well as GPI anchoring.

PMID: 20040362 [PubMed - as supplied by publisher]

Read the complete post at http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?tmpl=NoSidebarfile&db=PubMed&cmd=Retrieve&list_uids=20040362&dopt=Abstract

Posted 31 Dec 2009 7:59 AM by pubmed: neuroblastoma
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