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J. Biol. Chem., Vol. 280, Issue 13, 12494-12502, April 1, 2005
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From the
Brain Research Institute, University of Zürich and Department of Biology, ETH Zürich, CH-8057 Zürich, Switzerland and the ¶Laboratoire de Signalisations Moléculaires et Neurodégénérescence, EA 3433, Université Louis Pasteur, Faculté de Médecine, 11 rue Humann, 67085 Strasbourg Cedex, France
Nogo-A, -B, and -C are generated from the Nogo/RTN-4 gene and share a highly conserved C-terminal domain. They lack an N-terminal signal sequence and are predominantly localized to the endoplasmic reticulum (ER). We found the N terminus of endogenous Nogo-A exposed on the surface of fibroblasts, DRG neurons, and myoblasts. Surface-expressed Nogo-A was also present on presynaptic terminals of the neuromuscular junction and on DRG neurons in vivo. Surface biotinylations confirmed the presence of all Nogo isoforms on the surface. To search for proteins that interact with Nogo-A and suggest a function for the large intracellular pool of Nogo-A, immunoprecipitations were performed. Surprisingly, the most predominant proteins that interact with Nogo-A are Nogo-B and Nogo-C as seen with radiolabeled lysates and as confirmed by Western blotting in multiple cell lines. Nogo-A, -B, and -C share a 180-amino acid C-terminal domain with two highly conserved hydrophobic stretches that could form a channel or transporter in the ER and/or on the cell surface.
Received for publication, October 18, 2004 , and in revised form, January 6, 2005.
* This work was supported by Grants from the Swiss National Science Foundation (31-63633.00), the NCCR "Neural Plasticity and Repair" of the Swiss National Science Foundation, the Spinal Cord Consortium of the Christopher Reeve Paralysis Foundation (Springfield, NJ), and the Transregio-Sonderforschungsbereich Konstanz-Zurich, and the EU NeuroNetwork Project. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed: Brain Research Institute, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland. Tel.: 41-1-635-3262; Fax: 41-1-635-3303; E-mail: dodd{at}hifo.unizh.ch.
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