Two-dimensional crystallization of the mouse serotonin 5-HT3Areceptor.
The mouse serotonin 5-HT3Areceptor is a homo-pentameric ligand-gated ion channel (pLGIC) mediating fast excitatory neurotransmission in the central nervous system. The molecular mechanism of ion permeation of 5-HT3Areceptors triggered by the neurotransmitter serotonin is not yet fully understood. The recent X-ray structure of the mouse serotonin 5-HT3Areceptor in complex with a stabilizing nanobody revealed for the first time the entire structure of a mammalian pLGIC in detergent. Structural information of the receptor in a lipid bilayer however is still limited primarily due to the lack of 2D crystals of the receptor in a lipid bilayer. Here we present our results on the formation and improvement of diffracting 2D crystals of the mouse 5-HT3Aby limited proteolysis and addition of conformational nanobodies.
Laboratory of Biomolecular Research, Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
Jan
J
Rheinberger
Theranyx, 163 Avenue de Luminy, F-13288 Marseille, France.
Ghérici
G
Hassaine
BioEM lab, Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland.
Mohamed
M
Chami
Center for Cellular Imaging and NanoAnalytics, Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland.
Henning
H
Stahlberg
Laboratory of Physical Chemistry of Polymers and Membranes, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Horst
H
Vogel
Laboratory of Biomolecular Research, Paul Scherrer Institute, CH-5232 Villigen, Switzerland. Electronic address: xiao.li@psi.ch.
Xiaodan
X
Li
D000818:Animals; D020285:Cryoelectron Microscopy; D003460:Crystallization; D003461:Crystallography; D021621:Imaging, Three-Dimensional; D051379:Mice; D008958:Models, Molecular; D008968:Molecular Conformation; D044406:Receptors, Serotonin, 5-HT3 / Q000737:chemistry* / Q000648:ultrastructure; D061905:Single-Domain Antibodies
D044406:Receptors, Serotonin, 5-HT3; D061905:Single-Domain Antibodies
2D crystals; 5-HT3 receptor; Electron crystallography; Nanobody; Pentameric ligand-gated ion channel