TY  - CHAP
A1  - Das, Hirakjyoti
A1  - Zografakis, Alexandros
A1  - Oeljeklaus, Silke
A1  - Warscheid, Bettina
T1  - Analysis of Yeast Peroxisomes via Spatial Proteomics
T1  - Analyse von Hefeperoxisomen durch Spatial Proteomik
T2  - Peroxisomes
N2  - 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 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.
KW  - peroxisome purification
KW  - mass spectrometry
KW  - label-free quantification
KW  - protein localization
KW  - spatial proteomics
KW  - Saccharomyces cerevisiae
KW  - differential centrifugation
KW  - density gradient centrifugation
KW  - organellar mapping
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-327532
PB  - Springer
ET  - accepted version
ER  -