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Originally published In Press as doi:10.1074/mcp.M400117-MCP200 on April 25, 2005.
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Molecular & Cellular Proteomics 4:945-957, 2005.
© 2005 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

"Subcellular Proteomics" of Neuromelanin Granules Isolated from the Human Brain*

Florian Tribl{ddagger},§,¶, Manfred Gerlach||, Katrin Marcus**, Esther Asan{ddagger}{ddagger}, Thomas Tatschner§§, Thomas Arzberger¶¶,||||, Helmut E. Meyer**, Gerhard Bringmann§ and Peter Riederer{ddagger}

From the {ddagger} Department of Clinical Neurochemistry, Clinic and Polyclinic for Psychiatry and Psychotherapy, and "The National Parkinson Foundation Research Laboratories," Miami, Florida, at the Bayerische Julius-Maximilians-Universität Würzburg, Füchsleinstrasse 15, 97080 Würzburg, || Department of Clinical Neurochemistry, Clinic and Polyclinic for Child and Adolescent Psychiatry and Psychotherapy, Bayerische Julius-Maximilians-Universität Würzburg, Füchsleinstrasse 15, 97080 Würzburg, ** Medical Proteome Center, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum, {ddagger}{ddagger} Institute of Anatomy and Cell Biology, Bayerische Julius-Maximilians-Universität Würzburg, Koellikerstrasse 6, 97970 Würzburg, §§ Institute of Forensic Medicine, Bayerische Julius-Maximilians-Universität Würzburg, Versbacher Strasse 3, 97078 Würzburg, ¶¶ Institute of Pathology, Bayerische Julius-Maximilians-Universität Würzburg, Josef-Schneider-Strasse 2, 97080 Würzburg, and § Institute of Organic Chemistry, Bayerische Julius-Maximilians-Universität Würzburg, Am Hubland, 97074 Würzburg, Germany

"Subcellular proteomics" is currently the most effective approach to characterize subcellular compartments. Based on the powerful combination of subcellular fractionation and protein identification by LC-MS/MS we were able for the first time to 1) isolate intact neuromelanin granules from the human brain and 2) establish the first protein profile of these granules. This compartment containing neuromelanin (NM) is primarily located in the primate’s substantia nigra, one of the main brain regions that severely degenerates in Parkinson disease. We used mechanic tissue disaggregation, discontinuous sucrose gradient centrifugation, cell disruption, and organelle separation to isolate NM granules from human substantia nigra. Using transmission electron microscopy we demonstrated that the morphological characteristics of the isolated NM granules are similar to those described in human brain tissue. Fundamentally we found numerous proteins definitely demonstrating a close relationship of NM-containing granules with lysosomes or lysosome-related organelles originating from the endosome-lysosome lineage. Intriguingly we further revealed the presence of endoplasmic reticulum-derived chaperones, especially the transmembrane protein calnexin, which recently has been located in lysosome-related melanosomes and has been suggested to be a melanogenic chaperone.

Recipient of the Ph.D. scholarship DOC from the Austrian Academy of Sciences. To whom correspondence should be addressed. Tel.: 49-931-201-77300; Fax: 49-931-201-77220; E-mail: florian.tribl{at}mail.uni-wuerzburg.de


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