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Analysis of Yeast Peroxisomes via Spatial Proteomics
Analyse von Hefeperoxisomen durch Spatial Proteomik
Please always quote using this URN: urn:nbn:de:bvb:20-opus-327532
- Peroxisomes are ubiquitous organelles with essential functions in numerous cellular processes such as lipid metabolism, detoxification of reactive oxygen species and signaling. Knowledge of the peroxisomal proteome including multi-localized proteins and, most importantly, changes of its composition induced by altering cellular conditions or impaired peroxisome biogenesis and function is of paramount importance for a holistic view on peroxisomes and their diverse functions in a cellular context. In this chapter, we provide a spatial proteomicsPeroxisomes are ubiquitous organelles with essential functions in numerous cellular processes such as lipid metabolism, detoxification of reactive oxygen species and signaling. Knowledge of the peroxisomal proteome including multi-localized proteins and, most importantly, changes of its composition induced by altering cellular conditions or impaired peroxisome biogenesis and function is of paramount importance for a holistic view on peroxisomes and their diverse functions in a cellular context. In this chapter, we provide a spatial proteomics protocol specifically tailored to the analysis of the peroxisomal proteome of baker's yeast that enables the definition of the peroxisomal proteome under distinct conditions and to monitor dynamic changes of the proteome including the relocation of individual proteins to a different cellular compartment. The protocol comprises subcellular fractionation by differential centrifugation followed by Nycodenz density gradient centrifugation of a crude peroxisomal fraction, quantitative mass spectrometric measurements of subcellular and density gradient fractions and advanced computational data analysis, resulting in the establishment of organellar maps on a global scale.…
Author: | Hirakjyoti Das, Alexandros Zografakis, Silke OeljeklausORCiD, Bettina WarscheidORCiD |
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URN: | urn:nbn:de:bvb:20-opus-327532 |
Document Type: | Book article / Book chapter |
Faculties: | Fakultät für Chemie und Pharmazie / Lehrstuhl für Biochemie |
Language: | English |
Parent Title (English): | Peroxisomes |
Year of Completion: | 2023 |
Edition: | accepted version |
Publisher: | Springer |
Schriftenreihe: | Methods in Molecular Biology; 2643 |
Pagenumber: | 13-31 |
DOI: | https://doi.org/10.1007/978-1-0716-3048-8_2 |
Sonstige beteiligte Institutionen: | CIBSS Centre for Integrative Biological Signalling Studies, University of Freiburg |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 572 Biochemie |
Tag: | Saccharomyces cerevisiae; density gradient centrifugation; differential centrifugation; label-free quantification; mass spectrometry; organellar mapping; peroxisome purification; protein localization; spatial proteomics |
Release Date: | 2023/09/25 |
EU-Project number / Contract (GA) number: | 812968 |
OpenAIRE: | OpenAIRE |
Licence (German): | Deutsches Urheberrecht |