AI Chat Paper
Note: Please note that the following content is generated by AMiner AI. SciOpen does not take any responsibility related to this content.
{{lang === 'zh_CN' ? '文章概述' : 'Summary'}}
{{lang === 'en_US' ? '中' : 'Eng'}}
Chat more with AI
PDF (386.2 KB)
Collect
Submit Manuscript AI Chat Paper
Show Outline
Outline
Show full outline
Hide outline
Outline
Show full outline
Hide outline
Protocol | Open Access

Enrichment of ER tubule-derived microsomes by differential centrifugation and immunoprecipitation

Yiduo Liu1,2Junjie Hu1,2( )Bing Yan1( )
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
University of Chinese Academy of Sciences, Beijing 101408, China
Show Author Information

Graphical Abstract

Abstract

The endoplasmic reticulum (ER) is an essential component of the endomembrane system in eukaryotes and plays a crucial role in protein and lipid synthesis, as well as the maintenance of calcium homeostasis. Morphologically, the ER is composed primarily of sheets and tubules. The tubular ER is composed of a network of tubular membrane structures, each with diameters ranging from 30 to 50 nanometers. In recent years, there has been in-depth research on the molecular mechanisms of membrane shaping and membrane fusion of the tubular ER. However, there is still limited understanding of the specific physiological functions of the tubular ER. Here, we report a protocol that combines differential centrifugation and immunoprecipitation to specifically enrich microsomes originating from the tubular ER in yeast. The ER tubule-derived microsomes can be further used for proteomic and lipidomic studies or other biochemical analyses.

References

 

Anwar K, Klemm RW, Condon A, Severin KN, Zhang M, Ghirlando R, Hu J, Rapoport TA, Prinz WA (2012) The dynamin-like GTPase Sey1p mediates homotypic ER fusion in S. cerevisiae. J Cell Biol 197(2): 209−217

 

Bian X, Klemm RW, Liu TY, Zhang M, Sun S, Sui X, Liu X, Rapoport TA, Hu J (2011) Structures of the atlastin GTPase provide insight into homotypic fusion of endoplasmic reticulum membranes. Proc Natl Acad Sci USA 108(10): 3976−3981

 

Griffing LR (2010) Networking in the endoplasmic reticulum. Biochem Soc Trans 38(3): 747−753

 

Guillen-Samander A, De Camilli P (2023) Endoplasmic reticulum membrane contact sites, lipid transport, and neurodegeneration. Cold Spring Harb Perspect Biol 15(4): a041257. https://doi.org/10.1101/cshperspect.a041257

 

Hu J, Prinz WA, Rapoport TA (2011) Weaving the web of ER tubules. Cell 147(6): 1226−1231

 

Hu J, Shibata Y, Voss C, Shemesh T, Li Z, Coughlin M, Kozlov MM, Rapoport TA, Prinz WA (2008) Membrane proteins of the endoplasmic reticulum induce high-curvature tubules. Science 319(5867): 1247−1250

 

Hu J, Shibata Y, Zhu PP, Voss C, Rismanchi N, Prinz WA, Rapoport TA, Blackstone C (2009) A class of dynamin-like GTPases involved in the generation of the tubular ER network. Cell 138(3): 549−561

 

Kriechbaumer V (2018) ER microsome preparation in Arabidopsis thaliana. Methods Mol Biol 1691: 117−123

 

Li J, Yan B, Si H, Peng X, Zhang SL, Hu J (2017) Atlastin regulates store-operated calcium entry for nerve growth factor-induced neurite outgrowth. Sci Rep 7: 43490. https://doi.org/10.1038/srep43490

 

Phillips MJ, Voeltz GK (2016) Structure and function of ER membrane contact sites with other organelles. Nat Rev Mol Cell Biol 17(2): 69−82

 

Salinas S, Proukakis C, Crosby A, Warner TT (2008) Hereditary spastic paraplegia: clinical features and pathogenetic mechanisms. Lancet Neurol 7(12): 1127−1138

 

Shibata Y, Voss C, Rist JM, Hu J, Rapoport TA, Prinz WA, Voeltz GK (2008) The reticulon and DP1/Yop1p proteins form immobile oligomers in the tubular endoplasmic reticulum. J Biol Chem 283(27): 18892−18904

 

Voeltz GK, Prinz WA, Shibata Y, Rist JM, Rapoport TA (2006) A class of membrane proteins shaping the tubular endoplasmic reticulum. Cell 124(3): 573−586

 

Wang X, Li S, Wang H, Shui W, Hu J (2017) Quantitative proteomics reveal proteins enriched in tubular endoplasmic reticulum of Saccharomyces cerevisiae. Elife 6: e23816. https://doi.org/10.7554/eLife.23816

 

Xiang Y, Lyu R, Hu J (2023) Oligomeric scaffolding for curvature generation by ER tubule-forming proteins. Nat Commun 14(1): 2617. https://doi.org/10.1038/s41467-023-38294-y

 

Yan L, Sun S, Wang W, Shi J, Hu X, Wang S, Su D, Rao Z, Hu J, Lou Z (2015) Structures of the yeast dynamin-like GTPase Sey1p provide insight into homotypic ER fusion. J Cell Biol 210(6): 961−972

 

Zhang M, Wu F, Shi J, Zhu Y, Zhu Z, Gong Q, Hu J (2013) ROOT HAIR DEFECTIVE3 family of dynamin-like GTPases mediates homotypic endoplasmic reticulum fusion and is essential for Arabidopsis development. Plant Physiol 163(2): 713−720

Biophysics Reports
Pages 61-66
Cite this article:
Liu Y, Hu J, Yan B. Enrichment of ER tubule-derived microsomes by differential centrifugation and immunoprecipitation. Biophysics Reports, 2024, 10(2): 61-66. https://doi.org/10.52601/bpr.2023.230031

113

Views

6

Downloads

0

Crossref

0

Scopus

0

CSCD

Altmetrics

Received: 30 October 2023
Accepted: 15 December 2023
Published: 30 April 2024
© The Author(s) 2024

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Return