Dynamics and retention of misfolded proteins in native ER membranes

Sarah Nehls, Erik L. Snapp, Nelson B. Cole, Kristien J M Zaal, Anne K. Kenworthy, Theresa H. Roberts, Jan Ellenberg, John F. Presley, Eric Siggia, Jennifer Lippincott-Schwartz

Research output: Contribution to journalArticle

202 Citations (Scopus)

Abstract

When co-translationally inserted into endoplasmic reticulum (ER) membranes, newly synthesized proteins encounter the lumenal environment of the ER, which contains chaperone proteins that facilitate the folding reactions necessary for protein oligomerization, maturation and export from the ER. Here we show, using a temperature-sensitive variant of vesicular stomatitis virus G protein tagged with green fluorescent protein (VSVG-GFP), and fluorescence recovery after photobleaching (FRAP), the dynamics of association of folded and misfolded VSVG complexes with ER chaperones. We also investigate the potential mechanisms underlying protein retention in the ER. Misfolded VSVG-GFP complexes at 40 °C are highly mobile in ER membranes and do not reside in post-ER compartments, indicating that they are not retained in the ER by immobilization or retrieval mechanisms. These complexes are immobilized in ATP-depleted or tunicamycin-treated cells, in which VSVG-chaperone interactions are no longer dynamic. These results provide insight into the mechanisms of protein retention in the ER and the dynamics of protein-folding complexes in native ER membranes.

Original languageEnglish (US)
Pages (from-to)288-295
Number of pages8
JournalNature Cell Biology
Volume2
Issue number5
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Endoplasmic Reticulum
Membranes
Proteins
Protein Folding
Green Fluorescent Proteins
Fluorescence Recovery After Photobleaching
Tunicamycin
Immobilization
Adenosine Triphosphate
Temperature

ASJC Scopus subject areas

  • Cell Biology

Cite this

Nehls, S., Snapp, E. L., Cole, N. B., Zaal, K. J. M., Kenworthy, A. K., Roberts, T. H., ... Lippincott-Schwartz, J. (2000). Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology, 2(5), 288-295. https://doi.org/10.1038/35010558

Dynamics and retention of misfolded proteins in native ER membranes. / Nehls, Sarah; Snapp, Erik L.; Cole, Nelson B.; Zaal, Kristien J M; Kenworthy, Anne K.; Roberts, Theresa H.; Ellenberg, Jan; Presley, John F.; Siggia, Eric; Lippincott-Schwartz, Jennifer.

In: Nature Cell Biology, Vol. 2, No. 5, 2000, p. 288-295.

Research output: Contribution to journalArticle

Nehls, S, Snapp, EL, Cole, NB, Zaal, KJM, Kenworthy, AK, Roberts, TH, Ellenberg, J, Presley, JF, Siggia, E & Lippincott-Schwartz, J 2000, 'Dynamics and retention of misfolded proteins in native ER membranes', Nature Cell Biology, vol. 2, no. 5, pp. 288-295. https://doi.org/10.1038/35010558
Nehls S, Snapp EL, Cole NB, Zaal KJM, Kenworthy AK, Roberts TH et al. Dynamics and retention of misfolded proteins in native ER membranes. Nature Cell Biology. 2000;2(5):288-295. https://doi.org/10.1038/35010558
Nehls, Sarah ; Snapp, Erik L. ; Cole, Nelson B. ; Zaal, Kristien J M ; Kenworthy, Anne K. ; Roberts, Theresa H. ; Ellenberg, Jan ; Presley, John F. ; Siggia, Eric ; Lippincott-Schwartz, Jennifer. / Dynamics and retention of misfolded proteins in native ER membranes. In: Nature Cell Biology. 2000 ; Vol. 2, No. 5. pp. 288-295.
@article{5c529e1e8c7344e78ca3730b5bcf86d1,
title = "Dynamics and retention of misfolded proteins in native ER membranes",
abstract = "When co-translationally inserted into endoplasmic reticulum (ER) membranes, newly synthesized proteins encounter the lumenal environment of the ER, which contains chaperone proteins that facilitate the folding reactions necessary for protein oligomerization, maturation and export from the ER. Here we show, using a temperature-sensitive variant of vesicular stomatitis virus G protein tagged with green fluorescent protein (VSVG-GFP), and fluorescence recovery after photobleaching (FRAP), the dynamics of association of folded and misfolded VSVG complexes with ER chaperones. We also investigate the potential mechanisms underlying protein retention in the ER. Misfolded VSVG-GFP complexes at 40 °C are highly mobile in ER membranes and do not reside in post-ER compartments, indicating that they are not retained in the ER by immobilization or retrieval mechanisms. These complexes are immobilized in ATP-depleted or tunicamycin-treated cells, in which VSVG-chaperone interactions are no longer dynamic. These results provide insight into the mechanisms of protein retention in the ER and the dynamics of protein-folding complexes in native ER membranes.",
author = "Sarah Nehls and Snapp, {Erik L.} and Cole, {Nelson B.} and Zaal, {Kristien J M} and Kenworthy, {Anne K.} and Roberts, {Theresa H.} and Jan Ellenberg and Presley, {John F.} and Eric Siggia and Jennifer Lippincott-Schwartz",
year = "2000",
doi = "10.1038/35010558",
language = "English (US)",
volume = "2",
pages = "288--295",
journal = "Nature Cell Biology",
issn = "1465-7392",
publisher = "Nature Publishing Group",
number = "5",

}

TY - JOUR

T1 - Dynamics and retention of misfolded proteins in native ER membranes

AU - Nehls, Sarah

AU - Snapp, Erik L.

AU - Cole, Nelson B.

AU - Zaal, Kristien J M

AU - Kenworthy, Anne K.

AU - Roberts, Theresa H.

AU - Ellenberg, Jan

AU - Presley, John F.

AU - Siggia, Eric

AU - Lippincott-Schwartz, Jennifer

PY - 2000

Y1 - 2000

N2 - When co-translationally inserted into endoplasmic reticulum (ER) membranes, newly synthesized proteins encounter the lumenal environment of the ER, which contains chaperone proteins that facilitate the folding reactions necessary for protein oligomerization, maturation and export from the ER. Here we show, using a temperature-sensitive variant of vesicular stomatitis virus G protein tagged with green fluorescent protein (VSVG-GFP), and fluorescence recovery after photobleaching (FRAP), the dynamics of association of folded and misfolded VSVG complexes with ER chaperones. We also investigate the potential mechanisms underlying protein retention in the ER. Misfolded VSVG-GFP complexes at 40 °C are highly mobile in ER membranes and do not reside in post-ER compartments, indicating that they are not retained in the ER by immobilization or retrieval mechanisms. These complexes are immobilized in ATP-depleted or tunicamycin-treated cells, in which VSVG-chaperone interactions are no longer dynamic. These results provide insight into the mechanisms of protein retention in the ER and the dynamics of protein-folding complexes in native ER membranes.

AB - When co-translationally inserted into endoplasmic reticulum (ER) membranes, newly synthesized proteins encounter the lumenal environment of the ER, which contains chaperone proteins that facilitate the folding reactions necessary for protein oligomerization, maturation and export from the ER. Here we show, using a temperature-sensitive variant of vesicular stomatitis virus G protein tagged with green fluorescent protein (VSVG-GFP), and fluorescence recovery after photobleaching (FRAP), the dynamics of association of folded and misfolded VSVG complexes with ER chaperones. We also investigate the potential mechanisms underlying protein retention in the ER. Misfolded VSVG-GFP complexes at 40 °C are highly mobile in ER membranes and do not reside in post-ER compartments, indicating that they are not retained in the ER by immobilization or retrieval mechanisms. These complexes are immobilized in ATP-depleted or tunicamycin-treated cells, in which VSVG-chaperone interactions are no longer dynamic. These results provide insight into the mechanisms of protein retention in the ER and the dynamics of protein-folding complexes in native ER membranes.

UR - http://www.scopus.com/inward/record.url?scp=0033782030&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033782030&partnerID=8YFLogxK

U2 - 10.1038/35010558

DO - 10.1038/35010558

M3 - Article

C2 - 10806480

AN - SCOPUS:0033782030

VL - 2

SP - 288

EP - 295

JO - Nature Cell Biology

JF - Nature Cell Biology

SN - 1465-7392

IS - 5

ER -