title: Proteomic analysis of mouse astrocytes and their secretome by a combination of FASP and StageTip-based, high pH, reversed-phase fractionation.
abstract: Astrocytes are the most abundant cells in the CNS, but their function remains largely unknown. Characterization of the whole-cell proteome and secretome in astrocytes would facilitate the study of their functions in various neurodegenerative diseases and astrocyte-neuron communication. To build a reference proteome, we established a C8-D1A astrocyte proteome to a depth of 7265 unique protein groups using a novel strategy that combined two-step digestion, filter-aided sample preparation, StageTip-based high pH fractionation, and high-resolution MS. Nearly, 6000 unique protein groups were identified from conditioned media of astrocyte cultures, constituting the largest astrocyte secretome that has been reported. High-confidence whole-cell proteomes and secretomes are valuable resources in studying astrocyte function by label-free quantitation and bioinformatics analysis. All MS data have been deposited in the ProteomeXchange with identifier PXD000501 (http://proteomecentral.proteomexchange.org/dataset/PXD000501).
authors.affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea; Department of Biomedical Engineering, Seoul National University College of Medicine, Seoul, Korea; Institute of Medical & Biological Engineering, Medical Research Center, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Dohyun
authors.initials: D
authors.lastname: Han
authors.forename: Jonghwa
authors.initials: J
authors.lastname: Jin
authors.forename: Jongmin
authors.initials: J
authors.lastname: Woo
authors.forename: Hophil
authors.initials: H
authors.lastname: Min
authors.forename: Youngsoo
authors.initials: Y
authors.lastname: Kim
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000737:chemistry* / Q000378:metabolism; D002460:Cell Line; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D020543:Proteome
keywords: Astrocyte; High pH fractionation; LC-MS/MS; Proteome profile; Secretome; Technology