A nondesensitizing kainate receptor point mutant.
Ionotropic glutamate receptor (iGluR) desensitization can be modulated by mutations that change the stability of a dimer formed by the agonist binding domain. Desensitization of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors can be blocked by a single point mutation (e.g., GluR2 L483Y) that stabilizes this dimer in an active conformation. In contrast, desensitization of kainate receptors can be slowed, but not blocked, by similar dimer interface mutations. Only covalent cross-linking via introduced disulfides has been previously shown to block kainate receptor desensitization completely. We have now identified an apparently nondesensitizing GluR6 point mutant (D776K) located at the apex of the ligand binding (S1S2) domain dimer interface. Asp776 is one of a cluster of four charged residues in this region that together mediate direct dimer interactions and contribute to the binding sites for one chloride and two sodium ions. Despite the localized +4 change in the net charge of the S1S2 dimer, the D776K mutation actually increased the thermodynamic stability of the dimer. Unlike GluR6 wild type, the D776K mutant is insensitive to external cations but retains sensitivity to external anions. We therefore hypothesize that the unexpected phenotype of this charge reversal mutation results from the substitution of the sodium ions bound within the dimer interface by the introduced lysine NH(3)(+) groups. The nondesensitizing D776K mutant provides insights into kainate receptor gating and represents a potentially useful new tool for dissecting kainate receptor function.
Nayeem
Naushaba
N
Department of Pharmacology, School of Biomedical Sciences, University of Liverpool, Ashton Street, Liverpool L69 3GE, UK.
Zhang
Yihong
Y
Schweppe
Devin K
DK
Madden
Dean R
DR
Green
Tim
T
D019943:Amino Acid Substitution; D000818:Animals; D000838:Anions / Q000378:metabolism; D002414:Cations, Monovalent / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D006801:Humans; D017354:Point Mutation; D055503:Protein Multimerization / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D051381:Rats; D018092:Kainic Acid Receptors / Q000235:genetics* / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D000097806:GluK2 Kainate Receptor
D000838:Anions; D002414:Cations, Monovalent; D002712:Chlorides; D018092:Kainic Acid Receptors; D012964:Sodium; D000097806:GluK2 Kainate Receptor