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Published August 7, 2009 | Supplemental Material + Published
Journal Article Open

Monoclonal Antibodies Recognize Distinct Conformational Epitopes Formed by Polyglutamine in a Mutant Huntingtin Fragment

Abstract

Huntington disease (HD) is a neurodegenerative disorder caused by an expansion of a polyglutamine (polyQ) domain in the N-terminal region of huntingtin (htt). PolyQ expansion above 35–40 results in disease associated with htt aggregation into inclusion bodies. It has been hypothesized that expanded polyQ domains adopt multiple potentially toxic conformations that belong to different aggregation pathways. Here, we used atomic force microscopy to analyze the effect of a panel of anti-htt antibodies (MW1–MW5, MW7, MW8, and 3B5H10) on aggregate formation and the stability of a mutant htt-exon1 fragment. Two antibodies, MW7 (polyproline-specific) and 3B5H10 (polyQ-specific), completely inhibited fibril formation and disaggregated preformed fibrils, whereas other polyQ-specific antibodies had widely varying effects on aggregation. These results suggest that expanded polyQ domains adopt multiple conformations in solution that can be readily distinguished by monoclonal antibodies, which has important implications for understanding the structural basis for polyQ toxicity and the development of intrabody-based therapeutics for HD.

Additional Information

Copyright © 2009 by the American Society for Biochemistry and Molecular Biology. Received for publication, March 24, 2009 , and in revised form, May 4, 2009. Originally published In Press as doi:10.1074/jbc.M109.016923 on June 2, 2009 Supported by a postdoctoral fellowship from the Hereditary Disease Foundation. Supported by the National Institutes of Health-NIGMS UCSF Medical Scientist Training Program and a fellowship from the University of California at San Francisco Hillblom Center for the Biology of Aging. This work was supported, in whole or in part, by National Institutes of Health Grants R01NS047237 and R01NS054753 (to P. J. M.), P01AG022074 (to S. F.), R01NS039074 (to S. F.), and R01NS045091 and R01NS055298 (to P. H. P.). This work was also supported by the Hereditary Disease Foundation and the Cure Huntington's Disease Initiative. The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. 1 and Movies S1 and S2. We acknowledge Carl Johnson for insightful discussions and Gary Howard for editorial assistance.

Attached Files

Published - Legleiter2009p5619Journal_of_Biological_Chemistry.pdf

Supplemental Material - 1.pdf

Supplemental Material - Supplement_Fig.tif

Supplemental Material - jbc.M109.016923-1.mov

Supplemental Material - jbc.M109.016923-2.mov

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