An optimized quantitative proteomics method establishes the cell type-resolved mouse brain secretome
| Autor: | Johanna Tüshaus, Dmitrij Frishman, Minhui Su, Gökhan Güner, Stefan F. Lichtenthaler, Mikael Simons, Georg Jocher, Sabina Tahirovic, Jan Zaucha, Laura Sebastian Monasor, Stephan A. Müller, Evans Kataka |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Lipopolysaccharides
Proteomics metabolism [Antigens CD] Proteome Cell antagonists & inhibitors [Amyloid Precursor Protein Secretases] metabolism [Microglia] Mice 0302 clinical medicine Tandem Mass Spectrometry brain cells Aspartic Acid Endopeptidases secretomics cerebrospinal fluid [ADAM Proteins] Cells Cultured Neurons 0303 health sciences Principal Component Analysis metabolism [Astrocytes] General Neuroscience Brain Cerebrospinal Fluid Proteins BACE1 Secretomics antagonists & inhibitors [Aspartic Acid Endopeptidases] metabolism [Aspartic Acid Endopeptidases] ddc Cell biology pharmacology [Lipopolysaccharides] Oligodendroglia medicine.anatomical_structure cerebrospinal fluid [Aspartic Acid Endopeptidases] metabolism [Neurons] Microglia Resource Cell type Quantitative proteomics Nerve Tissue Proteins CSF Biology General Biochemistry Genetics and Molecular Biology cerebrospinal fluid [Antigens CD] metabolism [Oligodendroglia] 03 medical and health sciences proteomics Antigens CD cerebrospinal fluid [Amyloid Precursor Protein Secretases] ddc:570 medicine Animals Molecular Biology Neuroinflammation 030304 developmental biology metabolism [Nerve Tissue Proteins] General Immunology and Microbiology cytology [Brain] cerebrospinal fluid [Nerve Tissue Proteins] metabolism [Proteome] metabolism [Amyloid Precursor Protein Secretases] Mice Inbred C57BL ADAM Proteins Secretory protein Gene Ontology Membrane protein metabolism [Brain] Astrocytes metabolism [ADAM Proteins] Amyloid Precursor Protein Secretases methods [Proteomics] 030217 neurology & neurosurgery Software Neuroscience Chromatography Liquid |
| Zdroj: | The EMBO journal 39(20), e105693 (2020). doi:10.15252/embj.2020105693 The EMBO Journal |
| DOI: | 10.15252/embj.2020105693 |
| Popis: | To understand how cells communicate in the nervous system, it is essential to define their secretome, which is challenging for primary cells because of large cell numbers being required. Here, we miniaturized secretome analysis by developing the “high‐performance secretome protein enrichment with click sugars” (hiSPECS) method. To demonstrate its broad utility, hiSPECS was used to identify the secretory response of brain slices upon LPS‐induced neuroinflammation and to establish the cell type‐resolved mouse brain secretome resource using primary astrocytes, microglia, neurons, and oligodendrocytes. This resource allowed mapping the cellular origin of CSF proteins and revealed that an unexpectedly high number of secreted proteins in vitro and in vivo are proteolytically cleaved membrane protein ectodomains. Two examples are neuronally secreted ADAM22 and CD200, which we identified as substrates of the Alzheimer‐linked protease BACE1. hiSPECS and the brain secretome resource can be widely exploited to systematically study protein secretion and brain function and to identify cell type‐specific biomarkers for CNS diseases. hiSPECS, a miniaturized proteomics protocol based on pull‐down of glycosylated secretory proteins from smaller numbers of cells, defines the specific secretomes of astrocytes, microglia, neurons and oligodendrocytes from primary cells, as well as secretion changes in LPS‐induced inflammatory conditions. |
| Databáze: | OpenAIRE |
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