Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4

Shanti Pal Gangwar; Marriah N Green; Maria V Yelshanskaya; Alexander I Sobolevsky

doi:10.1016/j.xpro.2021.100855

Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4

STAR Protoc. 2021 Oct 1;2(4):100855. doi: 10.1016/j.xpro.2021.100855. eCollection 2021 Dec 17.

Authors

Shanti Pal Gangwar¹, Marriah N Green^{1

2}, Maria V Yelshanskaya¹, Alexander I Sobolevsky¹

Affiliations

¹ Department of Biochemistry and Molecular Biophysics, Columbia University, 650 West 168 Street, New York, NY 10032, USA.
² Institute of Human Nutrition, Columbia University, 630 West 168 Street, New York, NY 10032, USA.

Abstract

Ionotropic glutamate receptors (iGluRs) are ligand-gated ion channels that play crucial roles in the central nervous system. iGluR homologs, termed glutamate receptor-like channels (GLRs), have been found in plants. Investigating the structural and functional relationship between iGluRs and GLRs was limited by GLR protein expression, purification, and structural characterization. Here, we provide a detailed protocol for Arabidopsis thaliana GLR3.4 (AtGLR3.4) expression in a mammalian cell line and purification for structure determination by cryogenic electron microscopy (cryo-EM). For the complete details on the use and execution of this protocol, please refer to Green et al. (2021).

Keywords: Cryo-EM; Model Organisms; Plant sciences; Protein Biochemistry; Protein expression and purification.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Arabidopsis Proteins* / genetics
Arabidopsis* / genetics
Cryoelectron Microscopy
Mammals / metabolism
Receptors, Glutamate / genetics

Substances

Arabidopsis Proteins
Receptors, Glutamate

Grants and funding

R01 NS107253/NS/NINDS NIH HHS/United States