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PMID: 16029166 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The epididymal soluble prion protein forms a high-molecular-mass complex in association with hydrophobic proteins.

The Biochemical journal ·Vol. 392 ·No. Pt 1 ·2005-11-15 ·Pages 211-9

Ecroyd H, Belghazi M, Dacheux JL, Gatti JL

Abstract

We have shown previously that a 'soluble' form of PrP (prion protein), not associated with membranous vesicles, exists in the male reproductive fluid [Ecroyd, Sarradin, Dacheux and Gatti (2004) Biol. Reprod. 71, 993-1001]. Attempts to purify this 'soluble' PrP indicated that it behaves like a high-molecular-mass complex of more than 350 kDa and always co-purified with the same set of proteins. The main associated proteins were sequenced by MS and were found to match to clusterin (apolipoprotein J), BPI (bacterial permeability-increasing protein), carboxylesterase-like urinary excreted protein (cauxin), beta-mannosidase and beta-galactosidase. Immunoblotting and enzymatic assay confirmed the presence of clusterin and a cauxin-like protein and showed that a 17 kDa hydrophobic epididymal protein was also associated with this complex. These associated proteins were not separated by a high ionic strength treatment but were by 2-mercaptoethanol, probably due to its action on reducing disulphide bonds that maintain the interaction of components of the complex. Our results suggest that the associated PrP retains its GPI (glycosylphosphatidylinositol) anchor, in contrast with brain-derived PrP, and that it is resistant to cleavage by phosphatidylinositol-specific phospholipase C. Based on these results, the identity of the associated proteins and the overall biochemical properties of this protein ensemble, we suggest that 'soluble' PrP can form protein complexes that are maintained by hydrophobic interactions, in a similar manner to lipoprotein vesicles or micellar complexes.

MeSH Terms
Amino Acid Sequence Animals Clusterin/metabolism Epididymis/metabolism Hydrophobic and Hydrophilic Interactions Male Molecular Sequence Data Molecular Weight Multiprotein Complexes/chemistry,metabolism Prions/chemistry,metabolism Protein Binding Sheep Solubility
Chemicals
Clusterin Multiprotein Complexes Prions
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ecroyd Heath
Gamète Male et Fertilité, Institut National de la Recherche Agronomique, INRA-Nouzilly, 37380 Monnaie, France.
Belghazi Maya
Dacheux Jean-Louis
Gatti Jean-Luc
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
1470-8728
DOI
10.1042/BJ20050459
Published
2005-11-15
Pages
211-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1317680
Subset
IM
External Links
PubMed source DOI: 10.1042/BJ20050459 PMC Full Text View journal details
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