Skip to main content
SLU publication database (SLUpub)

Abstract

We have compared the processing, turnover and release of bovine PrP (boPrP) in transfected baby hamster kidney (BHK) and mouse neuroblastoma (N2a) cells. In BHK cells, boPrP was subjected to two distinct proteolytic cleavage events, the first was mapped between K-121 and H-122 generating an N-terminal and a C-terminal PrP fragment. Transport block experiments, cell surface biotinylation and PIPLC analyses showed that the bulk of boPrP on the cell surface was the C-terminal fragment and indicated that the first cleavage of boPrP took place prior to or very soon after it appears at the cell surface. The second cleavage was situated at the extreme C-terminus of the boPrP GPI-anchored C-terminal fragment and as a result of this was shed into the medium rapidly. The kinetics, the migration in SIDS-PAGE of the released fragment and protease inhibition studies indicate that a proteolytic activity was responsible for the release of the boPrP fragment from its GPI-anchor. Both N- and C-terminal fragments of boPrP could be detected in the medium. Moreover, in normal bovine brain, a C-terminal fragment was identified, suggesting that simlar proteolytic processing events occur in vivo. In N2a cells, the majority of boPrP was Subjected to a more complete degradation process, and only trace amounts of full length boPrP was shed into cell culture medium in a process which also indicated a release by proteolytic cleavage. (c) 2005 Elsevier B.V. All rights reserved

Keywords

Bovine prion; Cleavage site; Shedding; Soluble PrP

Published in

Virus Research
2006, volume: 115, number: 1, pages: 43-55
Publisher: ELSEVIER SCIENCE BV

SLU Authors

  • Klingeborn, Mikael

    • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

  • Linne, Tommy

    • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

  • Simonsson, Magnus

    • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

UKÄ Subject classification

Food Science

Publication identifier

  • DOI: https://doi.org/10.1016/j.virusres.2005.07.004

Permanent link to this page (URI)

https://res.slu.se/id/publ/7622