Quantitative proteomics reveal proteins enriched in tubular endoplasmic reticulum of Saccharomyces cerevisiae

eLife

Haicheng Wang¹  Xinbo Wang²  Junjie Hu³  Shanshan Li⁴  Wenqing Shui⁴ 

[1] College of Life Sciences, Huazhong University of Science and Technology, Wuhan, China;Department of Genetics and Cell Biology, College of Life Sciences, Nankai University, Tianjin, China;National Laboratory of Biomacromolecules and CAS Center for Excellence in Biomacromolecules, Institue of Biophysics, Chinese Academy of Sciences, Beijing, China;iHuman Institute and School of Life Science and Technology, ShanghaiTech University, Shanghai, China;

关键词: endoplasmic reticulum;    tubular network;    membrane curvature;    organelle isolation;    quantitative proteomics;   

DOI  :  10.7554/eLife.23816

来源: DOAJ

【 摘 要 】

The tubular network is a critical part of the endoplasmic reticulum (ER). The network is shaped by the reticulons and REEPs/Yop1p that generate tubules by inducing high membrane curvature, and the dynamin-like GTPases atlastin and Sey1p/RHD3 that connect tubules via membrane fusion. However, the specific functions of this ER domain are not clear. Here, we isolated tubule-based microsomes from Saccharomyces cerevisiae via classical cell fractionation and detergent-free immunoprecipitation of Flag-tagged Yop1p, which specifically localizes to ER tubules. In quantitative comparisons of tubule-derived and total microsomes, we identified a total of 79 proteins that were enriched in the ER tubules, including known proteins that organize the tubular ER network. Functional categorization of the list of proteins revealed that the tubular ER network may be involved in membrane trafficking, lipid metabolism, organelle contact, and stress sensing. We propose that affinity isolation coupled with quantitative proteomics is a useful tool for investigating ER functions.

【 授权许可】

Unknown