Abstract
Spinal bulbar muscular atrophy (SBMA) (also known as Kennedy’s disease) is a motor degenerative disease caused by an amplification of the polyglutamine stretch at the N-terminus of the human androgen receptor (AR). Amplifications larger than 40 glutamine residues are thought to lead to the disease. A characteristic feature of this disease is a ligand-dependent misfolding and aggregation of the mutant receptor that lead to the death of motor neurons. Initially, large cytoplasmic and nuclear aggregates reaching sizes of 6 μm were thought to be the pathogenic agents. Later studies have suggested that oligomeric species with sizes of less than 1 μm that occur prior to the formation of the larger aggregates are the toxic agents. However, there have been disagreements regarding the shape of these oligomers, as most studies have been carried out with peptide fragments of the androgen receptor containing different lengths of polyglutamine stretch. We have isolated the wild-type AR with a polyglutamine stretch of 22 (ARQ22) and a mutant receptor with a stretch of 65 (ARQ65) using a baculovirus system and have analyzed the oligomeric structures formed by these receptors with atomic force microscopy. This method has allowed us to determine the conformations of the full-length wild-type and mutant AR and revealed the conformation of the mutant AR that causes SBMA.
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Acknowledgements
This work was supported in part by a research grant from the Fritz Thyssen Stiftung to A.C.B.C. Research grants were also awarded to T.J. for this work from the Association Française Contre les Myopathies and from KIT “YIG” funds. We acknowledge with thanks the technical help of Rebecca Seeger and Jutta Stober. We also thank Donald McDonnell for providing us with the wild-type AR baculovirus expression plasmid pDW 464 AR. The work described here was carried out partly with the support of the Karlsruhe Nano Micro Facility (KNMF, www.knmf.kit.edu), a Helmholtz Research Infrastructure at Karlsruhe Institute of Technology (KIT, www.kit.edu).
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Jochum, T., Cato, A.C.B. (2014). Analysis of the Conformation of the Androgen Receptor in Spinal Bulbar Muscular Atrophy by Atomic Force Microscopy. In: Castoria, G., Auricchio, F. (eds) Steroid Receptors. Methods in Molecular Biology, vol 1204. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-1346-6_17
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DOI: https://doi.org/10.1007/978-1-4939-1346-6_17
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