Wiese Sebastian, Gronemeyer Thomas, Ofman Rob, Kunze Markus, Grou Cláudia P, Almeida José A, Eisenacher Martin, Stephan Christian, Hayen Heiko, Schollenberger Lukas, Korosec Thomas, Waterham Hans R, Schliebs Wolfgang, Erdmann Ralf, Berger Johannes, Meyer Helmut E, Just Wilhelm, Azevedo Jorge E, Wanders Ronald J A, Warscheid Bettina
Medizinisches Proteom-Center, Ruhr-Universitaet Bochum, Universitaetsstrasse 150, 44780 Bochum, Germany.
Mol Cell Proteomics. 2007 Dec;6(12):2045-57. doi: 10.1074/mcp.M700169-MCP200. Epub 2007 Sep 2.
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.
过氧化物酶体是一种普遍存在的、由单层膜包裹的关键细胞器，在哺乳动物中执行多种代谢途径，如通过α-氧化和β-氧化进行脂肪酸降解、醚磷脂生物合成、活性氧代谢以及乙醛酸解毒。为了履行这一系列广泛的代谢功能，过氧化物酶体容纳了至少约50种不同的酶，这是目前已确定的数量。对过氧化物酶体的兴趣因人类一组遗传疾病的发现而被激发，这些疾病是由过氧化物酶体生物发生缺陷或特定过氧化物酶体酶或转运蛋白的活性不足引起的。尽管这项研究极大地增进了我们对过氧化物酶体及其在哺乳动物代谢中作用的理解，但仍需要更深入地了解过氧化物酶体的生物化学和功能，以扩展我们对这个低丰度但至关重要的细胞器的认识。在这项工作中，我们使用经典的亚细胞分级分离技术结合基于质谱的蛋白质组学方法来表征小鼠肾脏过氧化物酶体的蛋白质组。我们几乎鉴定出了所有已知参与过氧化物酶体代谢和生物发生的成分。此外，通过使用定量质谱筛选结合统计分析的蛋白质定位研究，我们鉴定出了15个新的过氧化物酶体候选蛋白。其中，我们通过免疫细胞化学进一步研究了5个候选蛋白，证实了它们在过氧化物酶体中的定位。通过这项共同努力，我们相信已经编制出了迄今为止最全面的哺乳动物过氧化物酶体蛋白质目录。
