The major long-range goal of this work is obtain high resolution X-ray crystallographic structures of nicotinic acetylcholine receptors (AChRs). This family includes nicotinic acetylcholine (AChR), glycine, gamma- aminobutyric acidA (GABAA), and 5-hydroxytryptamine (5-HT3) receptors. All are involved in signal transmission within the nervous system and have wide-ranging importance in human neurophysiology and neurological diseases. The AChRs are particularly important clinically because of their roles in myasthenia gravis, nicotine addiction, and neurodegenerative diseases. Protein chemistry, ligand binding studies, electrophysiology, and electron microscopy on native and cloned AChRs have combined to elucidate some aspects of the structure and function of these receptors. The crystallographic structures of the AChRs, however, has been elusive. The alpha7 AChR is an attractive candidate for X-ray crystallography of nicotinic receptors because it possesses the simplifying property of containing only the alpha7 type of subunit. The proposed training environment is well-suited to the multidisciplinary nature of this project. The sponsor, Jon Lindstrom is a leader in the biology of AChRs whose laboratory is especially skilled in heterologous expression and functional analysis of AChRs. Patrick loll, an X-ray crystallographer and a collaborator on the project, is experienced with the crystallography of membrane proteins. The methods of protein design, expression and structural analysis complement the previous training of the principle investigator in protein magnetic resonance spectroscopy. They will have general relevance to this entire family ion channels and will be the foundation for his long-range goal of studying the structural biology of ion channels of the nervous system.
|Person, Alexandra M; Bills, Kathy L; Liu, Hong et al. (2005) Extracellular domain nicotinic acetylcholine receptors formed by alpha4 and beta2 subunits. J Biol Chem 280:39990-40002|
|Wells, G B; Lin, L; Jeanclos, E M et al. (2001) Assembly and ligand binding properties of the water-soluble extracellular domains of the glutamate receptor 1 subunit. J Biol Chem 276:3031-6|
|Anand, R; Nelson, M E; Gerzanich, V et al. (1998) Determinants of channel gating located in the N-terminal extracellular domain of nicotinic alpha7 receptor. J Pharmacol Exp Ther 287:469-79|
|Wells, G B; Anand, R; Wang, F et al. (1998) Water-soluble nicotinic acetylcholine receptor formed by alpha7 subunit extracellular domains. J Biol Chem 273:964-73|
|Diethelm-Okita, B; Wells, G; Kuryatov, A et al. (1998) Biosynthetic and synthetic AChR sequences to study T cells in myasthenia gravis. Ann N Y Acad Sci 841:320-3|
|Lindstrom, J; Peng, X; Kuryatov, A et al. (1998) Molecular and antigenic structure of nicotinic acetylcholine receptors. Ann N Y Acad Sci 841:71-86|
|Diethelm-Okita, B; Wells, G B; Kuryatov, A et al. (1998) Response of CD4+ T cells from myasthenic patients and healthy subjects of biosynthetic and synthetic sequences of the nicotinic acetylcholine receptor. J Autoimmun 11:191-203|