Arabidopsis thaliana glutamate receptor ion channel function demonstrated by ion pore transplantation.
Ionotropic glutamate receptors (iGluRs) are ligand-gated cation channels that mediate fast excitatory neurotransmission in the mammalian central nervous system. In the model plant Arabidopsis thaliana, a large family of 20 genes encoding proteins that share similarities with animal iGluRs in sequence and predicted secondary structure has been discovered. Members of this family, termed AtGLRs (A. thaliana glutamate receptors), have been implicated in root development, ion transport, and several metabolic and signalling pathways. However, there is still no direct proof of ligand-gated ion channel function of any AtGLR subunit. We used a domain transplantation technique to directly test whether the putative ion pore domains of AtGLRs can conduct ions. To this end, we transplanted the ion pore domains of 17 AtGLR subunits into rat alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (GluR1) and kainate (GluR6) receptor subunits and tested the resulting chimaeras for ion channel function in the Xenopus oocyte expression system. We show that AtGLR1.1 and AtGLR1.4 have functional Na(+)-, K(+)-, and Ca(2+)-permeable ion pore domains. The properties of currents through the AtGLR1.1 ion pore match those of glutamate-activated currents, depolarisations, and glutamate-triggered Ca(2+) influxes observed in plant cells. We conclude that some AtGLRs have functional non-selective cation pores.
Tapken
Daniel
D
Department of Biochemistry I-Receptor Biochemistry, NC6/170, Ruhr University Bochum, Bochum, Germany.
Hollmann
Michael
M
D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015220:Calcium Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004594:Electrophysiology; D005260:Female; D066298:In Vitro Techniques; D007473:Ion Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021122:Protein Subunits; D051381:Rats; D018091:Receptors, AMPA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017470:Receptors, Glutamate / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018092:Kainic Acid Receptors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014982:Xenopus laevis; D000097806:GluK2 Kainate Receptor
D029681:Arabidopsis Proteins; D015220:Calcium Channels; D007473:Ion Channels; D021122:Protein Subunits; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; D018092:Kainic Acid Receptors; D011993:Recombinant Fusion Proteins; D000097806:GluK2 Kainate Receptor; C496964:GLR1.1 protein, Arabidopsis; C585553:GLR1.4 protein, Arabidopsis