title: Quantitative proteomics reveal proteins enriched in tubular endoplasmic reticulum ofSaccharomyces cerevisiae.
abstract: 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 fromSaccharomyces cerevisiaevia 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.
authors.affiliation: Department of Genetics and Cell Biology, College of Life Sciences, Nankai University, Tianjin, China.
authors.forename: Xinbo
authors.initials: X
authors.lastname: Wang
authors.affiliation: iHuman Institute and School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Li
authors.affiliation: College of Life Sciences, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Haicheng
authors.initials: H
authors.lastname: Wang
authors.affiliation: iHuman Institute and School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
authors.forename: Wenqing
authors.initials: W
authors.lastname: Shui
authors.affiliation: National Laboratory of Biomacromolecules and CAS Center for Excellence in Biomacromolecules, Institue of Biophysics, Chinese Academy of Sciences, Beijing, China.
authors.forename: Junjie
authors.identifier: 0000-0003-4712-2243
authors.initials: J
authors.lastname: Hu
mesh_terms: D002458:Cell Fractionation; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D047468:Immunoprecipitation; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis*
chemical_list: D029701:Saccharomyces cerevisiae Proteins
keywords: S. cerevisiae; biochemistry; cell biology; endoplasmic reticulum; membrane curvature; organelle isolation; quantitative proteomics; tubular network