Modified ligand-gated ion channels and methods of use
AssigneeHoward Hughes Medical Institute
InventorsScott Sternson, Peter Lee, Christopher Magnus
Life of this patent
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Abstract
This document relates to materials and methods for controlling ligand gated ion channel (LGIC) activity. For example, modified LGICs including at least one LGIC subunit having a modified ligand binding domain (LBD) and/or a modified ion pore domain (IPD) are provided. Also provided are exogenous LGIC ligands that can bind to and activate the modified LGIC, as well as methods of modulating ion transport across the membrane of a cell of a mammal, methods of modulating the excitability of a cell in a mammal, and methods of treating a mammal having a channelopathy.
Claims31
1-35. (canceled)
A nucleic acid encoding a modified ligand gated ion channel (LGIC) comprising at least one modified LGIC subunit, said modified LGIC subunit comprising:
- (a) an alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having:
- (i) at least 93 percent sequence identity to a sequence set forth in residues 23-224 of SEQ ID NO:1, residues 23-229 of SEQ ID NO:2, or residues 23-233 of SEQ ID NO:11;
- and
- (ii) an amino acid modification comprising from 1-6 amino acid substitutions at one or more of amino acid residues selected from the group consisting of 27, 41, 77, 79, 115, 131, 139, 141, 175, 210, 216, 217, and 219 as numbered in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:11;
- wherein the amino acid substitution at residue 27 is R27D;
- wherein the amino acid substitution at residue 41 is E41R;
- wherein the amino acid substitution at residue 77 is W77F, W77Y, or W77M;
- wherein the amino acid substitution at residue 79 is Q79A, Q79G, or Q79S;
- wherein the amino acid substitution at residue 115 is Y115F;
- wherein the amino acid substitution at residue 131 is L131G, L131A, L131M, L131N, L131Q, L131V, or L131F;
- wherein the amino acid substitution at residue 139 is Q139L or Q139G;
- wherein the amino acid substitution at residue 141 is L141F or L141P;
- wherein the amino acid substitution at residue 175 is G175K, G175A, G175F, G175H, G175M, G175R, G175S, or G175V;
- wherein the amino acid substitution at residue 210 is Y210F;
- wherein the amino acid substitution at residue 216 is P216I;
- wherein the amino acid substitution at residue 217 is Y217F;
- and wherein the amino acid substitution at residue 219 is D219A;
- and
- (b) a glycine receptor (GlyR) ion pore domain (IPD).
A nucleic acid encoding a modified ligand gated ion channel (LGIC) comprising at least one modified LGIC subunit, said modified LGIC subunit comprising:
- (a) a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having:
- (i) at least 93 percent sequence identity to the sequence set forth in residues 23-224 of SEQ ID NO:1, residues 23-229 of SEQ ID NO:2, or residues 23-233 of SEQ ID NO:11;
- and
- (ii) an L131G amino acid substitution as numbered in SEQ ID NOs:
- 1, 2, or 11;
- and
- (b) a glycine receptor (GlyR) ion pore domain (IPD).
A nucleic acid encoding a modified ligand gated ion channel (LGIC) subunit, said modified LGIC subunit comprising:
- (a) a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having:
- (i) at least 93 percent sequence identity to the sequence set forth in residues 23-224 of SEQ ID NO:1, residues 23-229 of SEQ ID NO:2, or residues 23-233 of SEQ ID NO:11;
- and
- (ii) an L131G amino acid substitution as numbered in SEQ ID NOs:
- 1, 2, or 11;
- and
- (b) a glycine receptor (GlyR) ion pore domain (IPD).
Dependencies are inferred from the claim text.