Hans R Waterham

Publications (2)11.49 Total impact

• Article: The proteome of human liver peroxisomes: identification of five new peroxisomal constituents by a label-free quantitative proteomics survey.

  Thomas Gronemeyer, Sebastian Wiese, Rob Ofman, Christian Bunse, Magdalena Pawlas, Heiko Hayen, Martin Eisenacher, Christian Stephan, Helmut E Meyer, Hans R Waterham, Ralf Erdmann, Ronald J Wanders, Bettina Warscheid
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  ABSTRACT: The peroxisome is a key organelle of low abundance that fulfils various functions essential for human cell metabolism. Severe genetic diseases in humans are caused by defects in peroxisome biogenesis or deficiencies in the function of single peroxisomal proteins. To improve our knowledge of this important cellular structure, we studied for the first time human liver peroxisomes by quantitative proteomics. Peroxisomes were isolated by differential and Nycodenz density gradient centrifugation. A label-free quantitative study of 314 proteins across the density gradient was accomplished using high resolution mass spectrometry. By pairing statistical data evaluation, cDNA cloning and colocalization studies, we report the association of five new proteins with human liver peroxisomes. Among these, isochorismatase domain containing 1 protein points to the existence of a new metabolic pathway and hydroxysteroid dehydrogenase like 2 protein is likely involved in the transport or β-oxidation of fatty acids in human peroxisomes. The detection of alcohol dehydrogenase 1A suggests the presence of an alternative alcohol-oxidizing system in hepatic peroxisomes. In addition, lactate dehydrogenase A and malate dehydrogenase 1 partially associate with human liver peroxisomes and enzyme activity profiles support the idea that NAD becomes regenerated during fatty acid β-oxidation by alternative shuttling processes in human peroxisomes involving lactate dehydrogenase and/or malate dehydrogenase. Taken together, our data represent a valuable resource for future studies of peroxisome biochemistry that will advance research of human peroxisomes in health and disease.
  PLoS ONE 01/2013; 8(2):e57395. · 4.09 Impact Factor
• Article: Proteomics characterization of mouse kidney peroxisomes by tandem mass spectrometry and protein correlation profiling.

  Sebastian Wiese, Thomas Gronemeyer, Rob Ofman, Markus Kunze, Cláudia P Grou, José A Almeida, Martin Eisenacher, Christian Stephan, Heiko Hayen, Lukas Schollenberger, Thomas Korosec, Hans R Waterham, Wolfgang Schliebs, Ralf Erdmann, Johannes Berger, Helmut E Meyer, Wilhelm Just, Jorge E Azevedo, Ronald J A Wanders, Bettina Warscheid
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  ABSTRACT: 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.
  Molecular &amp Cellular Proteomics 01/2008; 6(12):2045-57. · 7.40 Impact Factor