Towards structure determination of neurotoxin II bound to nicotinic acetylcholine receptor: a solid‐state NMR approach

FEBS Letters

Castellani, Federica¹  Bocharov, Eduard³  Schulga, Alexey A³  Weise, Christoph²  Oschkinat, Hartmut¹  Hucho, Ferdinand²  Arseniev, Alexander S³  Krabben, Ludwig¹  van Rossum, Barth-Jan¹ 

[1] Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Straße 10, D-13125 Berlin, Germany;Institut für Chemie-Biochemie, Freie Universität Berlin, Thielallee 63, D-14195 Berlin, Germany;Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia

关键词: Binding site;    Magic-angle spinning nuclear magnetic resonance spectroscopy;    α-Neurotoxin;    Nicotinic acetylcholine receptor;    Solid-state nuclear magnetic resonance spectroscopy;    Bgtx;    α-bungarotoxin;    MAS;    magic-angle spinning;    nAChR;    nicotinic acetylcholine receptor;    NTII;    neurotoxin II;    PDSD;    proton-driven spin diffusion;    SH3;    α-spectrin Src-homology 3 domain;   

DOI  :  10.1016/S0014-5793(04)00252-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Solid-state magic-angle spinning nuclear magnetic resonance (NMR) has sufficient resolving power for full assignment of resonances and structure determination of immobilised biological samples as was recently shown for a small microcrystalline protein. In this work, we show that highly resolved spectra may be obtained from a system composed of a receptor–toxin complex. The NMR sample used for our studies consists of a membrane preparation of the nicotinic acetylcholine receptor from the electric organ of Torpedo californica which was incubated with uniformly ¹³C-,¹⁵N-labelled neurotoxin II. Despite the large size of the ligand–receptor complex (>290 kDa) and the high lipid content of the sample, we were able to detect and identify residues from the ligand. The comparison with solution NMR data of the free toxin indicates that its overall structure is very similar when bound to the receptor, but significant changes were observed for one isoleucine.

【 授权许可】

Unknown