2024-03-29T13:38:48Z
https://easy.dans.knaw.nl/oai/
oai:easy.dans.knaw.nl:easy-dataset:156149
2022-11-19T01:03:47Z
E10000
Zimmermann, R
via Mendeley Data
Proteomics explains client specificity of the human Sec62/Sec63 complex in ER protein import
Data Archiving and Networked Services (DANS)
2019
Interdisciplinary sciences
Richard Zimmermann
2019-12-05T16:03:04.745+01:00
2019-12-05T16:03:04.745+01:00
Dataset
10.17632/6s5hn73jcv.1
urn:nbn:nl:ui:13-wx-0f2e
easy-dataset:156149
info:eu-repo/semantics/openAccess
License: http://creativecommons.org/licenses/by/4.0
In mammalian cells, one-third of all polypeptides are transported into/across the ER-membrane via the Sec61-channel. While the Sec61-complex facilitates translocation of all polypeptides with amino-terminal signal peptides (SP) or transmembrane helices, the Sec61-associated Sec62/Sec63-complex supports translocation of only a subset, i.e. in a substrate-specific manner. To characterize Sec62- and Sec63-dependent precursors, we combined siRNA-mediated Sec62- or Sec63-depletion in human cells, label-free quantitative proteomics, and differential expression analysis. Alternatively, their CRISPR/Cas9-mediated knock-out in HEK 293 cells was carried out. The results were validated by western blotting under similar steady-state conditions and after short-term over-expression and simultaneous proteasome inhibition, respectively. SP analysis of Sec62- and Sec63-clients revealed the distinguishing feature. Thus, Sec62/Sec63-complex acts as an sp-receptor on the ER-membrane’s cytosolic face for certain precursor polypeptides and triggers substrate-specific (and regulated) opening of the Sec61-channel by its interactions with Sec61α.