eprintid: 8552
rev_number: 8
eprint_status: archive
userid: 1
dir: disk0/00/00/85/52
datestamp: 2008-07-15 17:26:54
lastmod: 2014-08-19 11:06:25
status_changed: 2012-08-14 15:25:52
type: article
metadata_visibility: show
creators_name: Meyer, David I.
creators_name: Louvard, Daniel
creators_name: Dobberstein, Bernhard
title: Characterization of Molecules Involved in Protein Translocation Using a Specific Antibody
ispublished: pub
subjects: ddc-570
divisions: i-706000
abstract: The vectorial translocation of nascent proteins through the membrane of the rough endoplasmic reticulum has been shown to require a specific membrane-bound protein whose  cytoplasmic domain can be proteolytically cleaved and isolated as an active peptide of mol wt 60,000 (Meyer and Dobberstein, 1980, J. Cell Biol. 87:503-508).  Rabbit antibodies raised against this peptide were used to further characterize the membrane-bound molecule. Immunoprecipitation of solubilized, radiolabeled rough microsomal proteins yielded a single polypeptide of mol wt 72,000, representing the membrane-bound protein from which the 60,000-mol wt peptide was proteolytically derived . The antibody could also be used to remove exclusively the 60,000-mol wt peptide, and thus the translocation activity, from elastase digests tested in a reconstituted system. Moreover,immunoprecipitation of elastase extracts alkylated with [t4C] N-ethylmaleimide selected a single species of mol wt 60,000. Immunoprecipitation of in vivo radiolabeled proteins from the appropriate cell type yielded the 72,000-mol wt membrane protein irrespective of the duration of labeling, or if followed by a chase. Subsequent treatment with protease generated the 60,000-mol wt fragment. In addition, the antibody could be used to visualize reticular structures in intact cells which correspond to endoplasmic reticulum at the ultrastructural level . It is thus clear that one membrane component required in the vectorial translocation of nascent secretory (and membrane) proteins is a peptide of mol wt 72,000.
abstract_translated_lang: eng
date: 1982
date_type: published
id_scheme: DOI
id_number: 10.11588/heidok.00008552
schriftenreihe_cluster_id: sr-2
schriftenreihe_order: 020
ppn_swb: 1354034929
own_urn: urn:nbn:de:bsz:16-opus-85528
language: eng
bibsort: MEYERDAVIDCHARACTERI1982
full_text_status: public
publication: The Journal of Cell Biology
volume: 92
pagerange: 579-583
citation:   Meyer, David I. ; Louvard, Daniel ; Dobberstein, Bernhard  (1982) Characterization of Molecules Involved in Protein Translocation Using a Specific Antibody.  The Journal of Cell Biology, 92.  pp. 579-583.      
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/8552/1/Meyer_1982.JCB.pdf