|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 276, Issue 10, 7475-7483, March 9, 2001
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Department of Neurobiology, Harvard Medical School,
Boston, Massachusetts 02115
Rapsyn, a 43-kDa peripheral membrane protein of
skeletal muscle, is essential for clustering nicotinic acetylcholine
receptors (nAChR) in the postsynaptic membrane. Previous studies with
rapsyn NH2-terminal fragments fused to green
fluorescent protein, expressed in 293T cells along with nAChRs,
establish the following: Rapsyn-(1-90), containing the
myristoylated amino terminus and two tetratricopeptide repeats (TPRs),
was sufficient for self-association at the plasma membrane;
rapsyn-(1-287), containing seven TPRs, did not cluster nAChRs; whereas
rapsyn-(1-360), containing a coiled-coil domain (rapsyn-(298-331)), clustered nAChRs. To further analyze the role of
rapsyn structural domains in self-association and nAChR clustering, we
have characterized the clustering properties of additional rapsyn
mutants containing deletions and substitutions within the TPR and
coiled-coil domains. A mutant lacking the coiled-coil domain alone
(rapsyn-(
Role of Rapsyn Tetratricopeptide Repeat and Coiled-coil Domains
in Self-association and Nicotinic Acetylcholine Receptor
Clustering*
288-348)), failed to cluster nAChRs. Within the
coiled-coil domain neutralization of the charged side chains was
tolerated, while alanine substitutions of large hydrophobic residues
resulted in the loss of nAChR clustering. Rapsyn self-association requires at least two TPRs, as a single TPR (TPR1 or TPR2 alone) was
not sufficient. While TPRs 1 and 2 are sufficient for self-association, they are not necessary, as TPRs 3-7 also formed clusters similar to
wild-type rapsyn. Fragments containing TPRs co-localized with full-length rapsyn, while the expressed coiled-coil or RING-H2 domain
did not. These results are discussed in terms of a homology model of
rapsyn, based on the three-dimensional structure of the TPR domain of
protein phosphatase 5.
*
This work was supported in part by United States Public
Health Service Grants NS 19522 and NS 18458 and by a Howard Hughes Medical Institute Predoctoral Fellowship (to M. J. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The 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: Dept. of
Neurobiology, Harvard Medical School, 220 Longwood Ave., Boston, MA
02115. Tel.: 617-432-1728; Fax: 617-734-7557; E-mail: jonathan_
cohen@hms.harvard.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Milone, X. M. Shen, D. Selcen, K. Ohno, J. Brengman, S. T. Iannaccone, C. M. Harper, and A. G. Engel Myasthenic syndrome due to defects in rapsyn: Clinical and molecular findings in 39 patients Neurology, July 21, 2009; 73(3): 228 - 235. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Cossins, G. Burke, S. Maxwell, H. Spearman, S. Man, J. Kuks, A. Vincent, J. Palace, C. Fuhrer, and D. Beeson Diverse molecular mechanisms involved in AChR deficiency due to rapsyn mutations Brain, October 1, 2006; 129(10): 2773 - 2783. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. R. Senechal, C. Thaller, and J. L. Noebels ADPEAF mutations reduce levels of secreted LGI1, a putative tumor suppressor protein linked to epilepsy Hum. Mol. Genet., June 15, 2005; 14(12): 1613 - 1620. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. L Gervasio and W. D Phillips Increased ratio of rapsyn to ACh receptor stabilizes postsynaptic receptors at the mouse neuromuscular synapse J. Physiol., February 1, 2005; 562(3): 673 - 685. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Ono, G. Mandel, and P. Brehm Acetylcholine Receptors Direct Rapsyn Clusters to the Neuromuscular Synapse in Zebrafish J. Neurosci., June 16, 2004; 24(24): 5475 - 5481. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Taipale, N. Kaminen, J. Nopola-Hemmi, T. Haltia, B. Myllyluoma, H. Lyytinen, K. Muller, M. Kaaranen, P. J. Lindsberg, K. Hannula-Jouppi, et al. From the Cover: A candidate gene for developmental dyslexia encodes a nuclear tetratricopeptide repeat domain protein dynamically regulated in brain PNAS, September 30, 2003; 100(20): 11553 - 11558. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. E. Coyle and D. B. Nikolov GABARAP: Lessons for Synaptogenesis Neuroscientist, June 1, 2003; 9(3): 205 - 216. [Abstract] [PDF]
|
|
|
|
 |
|
 P Richard, K Gaudon, F Andreux, E Yasaki, C Prioleau, S Bauche, A Barois, C Ioos, M Mayer, M C Routon, et al. Possible founder effect of rapsyn N88K mutation and identification of novel rapsyn mutations in congenital myasthenic syndromes J. Med. Genet., June 1, 2003; 40(6): e81 - 81. [Full Text] [PDF]
|
|
|
|
 |
|
 N. Amada, T. Tezuka, A. Mayeda, K. Araki, N. Takei, K. Todokoro, and H. Nawa A Novel Rat Orthologue and Homologue for the Drosophila crooked neck Gene in Neural Stem Cells and Their Immediate Descendants J. Biochem., May 1, 2003; 133(5): 615 - 623. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Ohno, M. Sadeh, I. Blatt, J. M. Brengman, and A. G. Engel E-box mutations in the RAPSN promoter region in eight cases with congenital myasthenic syndrome Hum. Mol. Genet., April 1, 2003; 12(7): 739 - 748. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Marchand, A. Devillers-Thiery, S. Pons, J.-P. Changeux, and J. Cartaud Rapsyn Escorts the Nicotinic Acetylcholine Receptor Along the Exocytic Pathway via Association with Lipid Rafts J. Neurosci., October 15, 2002; 22(20): 8891 - 8901. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Ono, A. Shcherbatko, S.-i. Higashijima, G. Mandel, and P. Brehm The Zebrafish Motility Mutant twitch once Reveals New Roles for Rapsyn in Synaptic Function J. Neurosci., August 1, 2002; 22(15): 6491 - 6498. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Boudin and A. M. Craig Molecular Determinants for PICK1 Synaptic Aggregation and mGluR7a Receptor Coclustering. ROLE OF THE PDZ, COILED-COIL, AND ACIDIC DOMAINS J. Biol. Chem., August 3, 2001; 276(32): 30270 - 30276. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Bartoli, M. K. Ramarao, and J. B. Cohen Interactions of the Rapsyn RING-H2 Domain with Dystroglycan J. Biol. Chem., June 29, 2001; 276(27): 24911 - 24917. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |