NDLI: Electron microscopic evidence for nucleation and growth of 3D acetylcholine receptor microcrystals in structured lipid

Content Provider	PubMed Central
Author	Paas, Yoav Jean, Cartaud Michel, Recouvreur Regis, Grailhe Dufresne, Virginie Eva, Pebay-peyroula Landau, Ehud M. Changeux, Jean-pierre
Copyright Year	2003
Abstract	Nicotinic acetylcholine receptors (AChRs) belong to a superfamily of oligomeric proteins that transduce electric signals across the cell membrane on binding of neurotransmitters. These receptors harbor a large extracellular ligand-binding domain directly linked to an ion-conducting channel-forming domain that spans the cell membrane 20 times and considerably extends into the cytoplasm. Thus far, none of these receptor channels has been crystallized in three dimensions. The crystallization of the AChR from Torpedo marmorata electric organs is challenged here in lipidic–detergent matrices. Detergent-soluble AChR complexed with α-bungarotoxin (αBTx), a polypeptidic competitive antagonist, was purified. The AChR–αBTx complex was reconstituted in a lipidic matrix composed of monoolein bilayers that are structured in three dimensions. The αBTx was conjugated to a photo-stable fluorophore, enabling us to monitor the physical behavior of the receptor–toxin complex in the lipidic matrix under light stereomicroscope, and to freeze fracture regions containing the receptor–toxin complex for visualization under a transmission electron microscope. Conditions were established for forming 2D receptor–toxin lattices that are stacked in the third dimension. 3D AChR nanocrystals were thereby grown inside the highly viscous lipidic 3D matrix. Slow emulsification of the lipidic matrix converted these nanocrystals into 3D elongated thin crystal plates of micrometer size. The latter are stable in detergent-containing aqueous solutions and can currently be used for seeding and epitaxial growth, en route to crystals of appropriate dimensions for x-ray diffraction studies.
Related Links	http://dx.doi.org/10.1073/pnas.1834451100
Ending Page	11314
Page Count	6
Starting Page	11309
File Format	PDF
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	20
Volume Number	100
Language	English
Publisher	National Academy of Sciences
Publisher Date	2003-09-30
Access Restriction	Open
Rights Holder	National Academy of Sciences
Subject Keyword	General Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

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