Peroxisomes from the heavy mitochondrial fraction

• Verfasst von: Islinger, Markus [VerfasserIn]
• Verfasst von: Li, Ka Wan [VerfasserIn]
• Verfasst von: Loos, Maarten [VerfasserIn]
• Verfasst von: Liebler, Sven Stefan [VerfasserIn]
• Verfasst von: Angermüller, Sabine [VerfasserIn]
• Verfasst von: Eckerskorn, Christoph [VerfasserIn]
• Verfasst von: Weber, Gerhard [VerfasserIn]
• Verfasst von: Abdolzade, Afsaneh [VerfasserIn]
• Verfasst von: Völkl, Alfred [VerfasserIn]
• Titel: Peroxisomes from the heavy mitochondrial fraction
• Titelzusatz: isolation by zonal free flow electrophoresis and quantitative mass spectrometrical characterization
• Verf.angabe: Markus Islinger, Ka Wan Li, Maarten Loos, Sven Liebler, Sabine Angermüller, Christoph Eckerskorn, Gerhard Weber, Afsaneh Abdolzade, and Alfred Völkl
• E-Jahr: 2010
• Jahr: 4 January 2010
• Umfang: 12 S.
• Illustrationen: Illustrationen
• Fussnoten: Published online: 9 September 2009 ; Gesehen am 11.07.2023
• Titel Quelle: Enthalten in: Journal of proteome research
• Ort Quelle: Washington, DC : ACS Publications, 2002
• Jahr Quelle: 2010
• Band/Heft Quelle: 9(2010), 1 vom: Jan., Seite 113-124
• ISSN Quelle: 1535-3907
• DOI: doi:10.1021/pr9004663
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1852369426

Abstract

Peroxisomes are a heterogeneous group of organelles fulfilling reactions in a variety of metabolic pathways. To investigate if functionally different subpopulations can be found within a single tissue, peroxisomes from the heavy mitochondrial fraction (HM-Po) of the rat liver were isolated and compared to “classic” peroxisomes from the light mitochondrial fraction (LM-Po) using iTRAQ tandem mass spectrometry. Peroxisomes represent only a minor although significant proportion of the heavy mitochondrial fraction (2700gmax) precluding a straightforward isolation by standard protocols. Thus, a new fractionation scheme suitable for a subsequent mass spectrometrical analysis was developed using a combination of centrifugation techniques and zonal free flow electrophoresis. On the basis of the iTRAQ-measurement, a variation of the peroxisomal protein pattern between both fractions could be determined and further confirmed by immunoblotting and enzyme activity assays for selected proteins: whereas peroxisomes from the light mitochondrial fraction contain high amounts of β-oxidation enzymes, peroxisomes from the heavy mitochondrial fraction were dominated by enzymes fulfilling other functions. Among other findings, HM-Po was characterized by a high abundance of D-amino acid oxidase. This observation can be mirrored at the ultrastructural level, where tissue sections of liver peroxisomes show a heterogeneous staining for the enzymes activity, when visualized by the cerium technique.