Proteomics characterization of mouse kidney peroxisomes by tandem mass spectrometry and protein correlation profiling.
The peroxisome represents a ubiquitous single membrane-bound key organelle that executes various metabolic pathways such as fatty acid degradation by alpha- and beta-oxidation, ether-phospholipid biosynthesis, metabolism of reactive oxygen species, and detoxification of glyoxylate in mammals. To fulfil this vast array of metabolic functions, peroxisomes accommodate approximately 50 different enzymes at least as identified until now. Interest in peroxisomes has been fueled by the discovery of a group of genetic diseases in humans, which are caused by either a defect in peroxisome biogenesis or the deficient activity of a distinct peroxisomal enzyme or transporter. Although this research has greatly improved our understanding of peroxisomes and their role in mammalian metabolism, deeper insight into biochemistry and functions of peroxisomes is required to expand our knowledge of this low abundance but vital organelle. In this work, we used classical subcellular fractionation in combination with MS-based proteomics methodologies to characterize the proteome of mouse kidney peroxisomes. We could identify virtually all known components involved in peroxisomal metabolism and biogenesis. Moreover through protein localization studies by using a quantitative MS screen combined with statistical analyses, we identified 15 new peroxisomal candidates. Of these, we further investigated five candidates by immunocytochemistry, which confirmed their localization in peroxisomes. As a result of this joint effort, we believe to have compiled the so far most comprehensive protein catalogue of mammalian peroxisomes.
Wiese
Sebastian
S
Medizinisches Proteom-Center, Ruhr-Universitaet Bochum, Universitaetsstrasse 150, 44780 Bochum, Germany.
Gronemeyer
Thomas
T
Ofman
Rob
R
Kunze
Markus
M
Grou
Cláudia P
CP
Almeida
José A
JA
Eisenacher
Martin
M
Stephan
Christian
C
Hayen
Heiko
H
Schollenberger
Lukas
L
Korosec
Thomas
T
Waterham
Hans R
HR
Schliebs
Wolfgang
W
Erdmann
Ralf
R
Berger
Johannes
J
Meyer
Helmut E
HE
Just
Wilhelm
W
Azevedo
Jorge E
JE
Wanders
Ronald J A
RJ
Warscheid
Bettina
B
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007668:Kidney / Q000378:metabolism*; D051379:Mice; D020675:Peroxisomes / Q000378:metabolism*; D040901:Proteomics; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*