 |
Previous Article | Next Article 
Volume 16, Number 17,
Issue of September 1, 1996
pp. 5415-5424
Copyright ©1996 Society for Neuroscience
Stoichiometry of a Recombinant GABAA Receptor
Received May 1, 1996; revised June 14, 1996; accepted June 18, 1996.
Yongchang Chang,
Ruoping Wang,
Sonal Barot, and
David S. Weiss
Neurobiology Research Center, Department of Physiology and
Biophysics, University of Alabama at Birmingham, Birmingham, Alabama
35294
GABA is the main inhibitory neurotransmitter in the mammalian
brain. The postsynaptic GABAA receptor/pore complex is
presumed to be a pentamer typically composed of a combination of ,
, and subunits, although the stoichiometry remains
controversial. We probed the stoichiometry of the GABAA
receptor by site-directed mutagenesis of a conserved leucine (to
serine) in the putative second membrane-spanning domain of the rat
1( L263S), 2( L259S), and 2( L274S) subunit isoforms.
Coexpression of wild-type and mutant subunits of each class (e.g., and L263S), along with their wild-type counterparts (e.g., and
), in Xenopus laevis oocytes resulted in mixed
populations of receptors with distinct GABA sensitivities. This is
consistent with the interpretation that the leucine mutation increased
the GABA sensitivity in proportion to the number of incorporated mutant
subunits. The apparent number of incorporated subunits for each class
( , , and ) could then be determined from the number of
components comprising the compound GABA dose-response relationships.
Using this approach, we conclude that the recombinant 1 2 2
GABAA receptor is a pentamer composed of two subunits,
two subunits, and one subunit.
Key words:
GABA;
stoichiometry;
receptor;
ion channel;
mutagenesis
This article has been cited by other articles:

|
 |

|
 |
 
B. E. Erkkila, A. V. Sedelnikova, and D. S. Weiss
Stoichiometric Pore Mutations of the GABAAR Reveal a Pattern of Hydrogen Bonding with Picrotoxin
Biophys. J.,
June 1, 2008;
94(11):
4299 - 4306.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. Mizuta, D. Xu, Y. Pan, G. Comas, J. R. Sonett, Y. Zhang, R. A. Panettieri Jr., J. Yang, and C. W. Emala Sr.
GABAA receptors are expressed and facilitate relaxation in airway smooth muscle
Am J Physiol Lung Cell Mol Physiol,
June 1, 2008;
294(6):
L1206 - L1216.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Keramidas and N. L. Harrison
Agonist-dependent Single Channel Current and Gating in {alpha}4{beta}2{delta} and {alpha}1{beta}2{gamma}2S GABAA Receptors
J. Gen. Physiol.,
January 28, 2008;
131(2):
163 - 181.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. R. Tan, A. Gonthier, R. Baur, M. Ernst, M. Goeldner, and E. Sigel
Proximity-accelerated Chemical Coupling Reaction in the Benzodiazepine-binding Site of {gamma}-Aminobutyric Acid Type A Receptors: SUPERPOSITION OF DIFFERENT ALLOSTERIC MODULATORS
J. Biol. Chem.,
September 7, 2007;
282(36):
26316 - 26325.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. H. Hadley and J. Amin
Rat {alpha}6beta2{delta} GABAA receptors exhibit two distinct and separable agonist affinities
J. Physiol.,
June 15, 2007;
581(3):
1001 - 1018.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. J. Yi and M. D. Ehlers
Emerging Roles for Ubiquitin and Protein Degradation in Neuronal Function
Pharmacol. Rev.,
March 1, 2007;
59(1):
14 - 39.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Zeller, M. Arras, R. Jurd, and U. Rudolph
Identification of a Molecular Target Mediating the General Anesthetic Actions of Pentobarbital
Mol. Pharmacol.,
March 1, 2007;
71(3):
852 - 859.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
I. Sarto-Jackson, R. Furtmueller, M. Ernst, S. Huck, and W. Sieghart
Spontaneous Cross-link of Mutated {alpha}1 Subunits during GABAA Receptor Assembly
J. Biol. Chem.,
February 16, 2007;
282(7):
4354 - 4363.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. J. Bailey, M. A. Ravier, and G. A. Rutter
Glucose-Dependent Regulation of {gamma}-Aminobutyric Acid (GABAA) Receptor Expression in Mouse Pancreatic Islet {alpha}-Cells
Diabetes,
February 1, 2007;
56(2):
320 - 327.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. R. McCartney, T. Z. Deeb, T. N. Henderson, and T. G. Hales
Tonically Active GABAA Receptors in Hippocampal Pyramidal Neurons Exhibit Constitutive GABA-Independent Gating
Mol. Pharmacol.,
February 1, 2007;
71(2):
539 - 548.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. H. Lagrange, E. J. Botzolakis, and R. L. Macdonald
Enhanced macroscopic desensitization shapes the response of {alpha}4 subtype-containing GABAA receptors to synaptic and extrasynaptic GABA
J. Physiol.,
February 1, 2007;
578(3):
655 - 676.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Sedelnikova, B. E. Erkkila, H. Harris, S. O. Zakharkin, and D. S. Weiss
Stoichiometry of a pore mutation that abolishes picrotoxin-mediated antagonism of the GABAA receptor
J. Physiol.,
December 1, 2006;
577(2):
569 - 577.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
Y. Chen, K. Reilly, and Y. Chang
Evolutionarily Conserved Allosteric Network in the Cys Loop Family of Ligand-gated Ion Channels Revealed by Statistical Covariance Analyses
J. Biol. Chem.,
June 30, 2006;
281(26):
18184 - 18192.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. Serantes, F. Arnalich, M. Figueroa, M. Salinas, E. Andres-Mateos, R. Codoceo, J. Renart, C. Matute, C. Cavada, A. Cuadrado, et al.
Interleukin-1beta Enhances GABAA Receptor Cell-surface Expression by a Phosphatidylinositol 3-Kinase/Akt Pathway: RELEVANCE TO SEPSIS-ASSOCIATED ENCEPHALOPATHY
J. Biol. Chem.,
May 26, 2006;
281(21):
14632 - 14643.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J B Vincent, S. Horike, S Choufani, A D Paterson, W Roberts, P Szatmari, R Weksberg, B Fernandez, and S W Scherer
An inversion inv(4)(p12-p15.3) in autistic siblings implicates the 4p GABA receptor gene cluster
J. Med. Genet.,
May 1, 2006;
43(5):
429 - 434.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. E. Petroski, J. E. Pomeroy, R. Das, H. Bowman, W. Yang, A. P. Chen, and A. C. Foster
Indiplon Is a High-Affinity Positive Allosteric Modulator with Selectivity for {alpha}1 Subunit-Containing GABAA Receptors
J. Pharmacol. Exp. Ther.,
April 1, 2006;
317(1):
369 - 377.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. M. McCann, J. Bracamontes, J. H. Steinbach, and J. R. Sanes
The cholinergic antagonist {alpha}-bungarotoxin also binds and blocks a subset of GABA receptors
PNAS,
March 28, 2006;
103(13):
5149 - 5154.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. M. White
Pretty Subunits All in a Row: Using Concatenated Subunit Constructs to Force the Expression of Receptors with Defined Subunit Stoichiometry and Spatial Arrangement
Mol. Pharmacol.,
February 1, 2006;
69(2):
407 - 410.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. J. Boileau, R. A. Pearce, and C. Czajkowski
Tandem Subunits Effectively Constrain GABAA Receptor Stoichiometry and Recapitulate Receptor Kinetics But Are Insensitive to GABAA Receptor-Associated Protein
J. Neurosci.,
December 7, 2005;
25(49):
11219 - 11230.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
P. V. Plazas, E. Katz, M. E. Gomez-Casati, C. Bouzat, and A. B. Elgoyhen
Stoichiometry of the {alpha}9{alpha}10 Nicotinic Cholinergic Receptor
J. Neurosci.,
November 23, 2005;
25(47):
10905 - 10912.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. K. Bera and M. H. Akabas
Spontaneous Thermal Motion of the GABAA Receptor M2 Channel-lining Segments
J. Biol. Chem.,
October 21, 2005;
280(42):
35506 - 35512.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. Baur, U. Simmen, M. Senn, U. Sequin, and E. Sigel
Novel Plant Substances Acting as {beta} Subunit Isoform-Selective Positive Allosteric Modulators of GABAA Receptors
Mol. Pharmacol.,
September 1, 2005;
68(3):
787 - 792.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D. M. Jones-Davis, L. Song, M. J. Gallagher, and R. L. Macdonald
Structural Determinants of Benzodiazepine Allosteric Regulation of GABAA Receptor Currents
J. Neurosci.,
August 31, 2005;
25(35):
8056 - 8065.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. Baur and E. Sigel
Benzodiazepines Affect Channel Opening of GABAA Receptors Induced by Either Agonist Binding Site
Mol. Pharmacol.,
April 1, 2005;
67(4):
1005 - 1008.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Sedelnikova, C. D. Smith, S. O. Zakharkin, D. Davis, D. S. Weiss, and Y. Chang
Mapping the {rho}1 GABAC Receptor Agonist Binding Pocket: CONSTRUCTING A COMPLETE MODEL
J. Biol. Chem.,
January 14, 2005;
280(2):
1535 - 1542.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. Jung, M. H. Akabas, and R. A. Harris
Functional and Structural Analysis of the GABAA Receptor {alpha}1 Subunit during Channel Gating and Alcohol Modulation
J. Biol. Chem.,
January 7, 2005;
280(1):
308 - 316.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. G. Newell, R. A. McDevitt, and C. Czajkowski
Mutation of Glutamate 155 of the GABAA Receptor {beta}2 Subunit Produces a Spontaneously Open Channel: A Trigger for Channel Activation
J. Neurosci.,
December 15, 2004;
24(50):
11226 - 11235.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
F. Sancar and C. Czajkowski
A GABAA Receptor Mutation Linked to Human Epilepsy ({gamma}2R43Q) Impairs Cell Surface Expression of {alpha}{beta}{gamma} Receptors
J. Biol. Chem.,
November 5, 2004;
279(45):
47034 - 47039.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. Kang and R. L. Macdonald
The GABAA Receptor {gamma}2 Subunit R43Q Mutation Linked to Childhood Absence Epilepsy and Febrile Seizures Causes Retention of {alpha}1{beta}2{gamma}2S Receptors in the Endoplasmic Reticulum
J. Neurosci.,
October 6, 2004;
24(40):
8672 - 8677.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
P. J. Groot-Kormelink, S. D. Broadbent, J. P. Boorman, and L. G. Sivilotti
Incomplete Incorporation of Tandem Subunits in Recombinant Neuronal Nicotinic Receptors
J. Gen. Physiol.,
June 1, 2004;
123(6):
697 - 708.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
B. C. Drafts and J. L. Fisher
Structural Determinants of the Pharmacological Properties of the GABAA Receptor {alpha}6 Subunit
J. Pharmacol. Exp. Ther.,
June 1, 2004;
309(3):
1108 - 1115.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
F. Minier and E. Sigel
Positioning of the {alpha}-subunit isoforms confers a functional signature to {gamma}-aminobutyric acid type A receptors
PNAS,
May 18, 2004;
101(20):
7769 - 7774.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D. Rusch, H. Zhong, and S. A. Forman
Gating Allosterism at a Single Class of Etomidate Sites on {alpha}1{beta}2{gamma}2L GABAA Receptors Accounts for Both Direct Activation and Agonist Modulation
J. Biol. Chem.,
May 14, 2004;
279(20):
20982 - 20992.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. Akk, J. Bracamontes, and J. H. Steinbach
Activation of GABAA receptors containing the {alpha}4 subunit by GABA and pentobarbital
J. Physiol.,
April 15, 2004;
556(2):
387 - 399.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
E. N. Goren, D. C. Reeves, and M. H. Akabas
Loose Protein Packing around the Extracellular Half of the GABAA Receptor {beta}1 Subunit M2 Channel-lining Segment
J. Biol. Chem.,
March 19, 2004;
279(12):
11198 - 11205.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D. Berezhnoy, Y. Nyfeler, A. Gonthier, H. Schwob, M. Goeldner, and E. Sigel
On the Benzodiazepine Binding Pocket in GABAA Receptors
J. Biol. Chem.,
January 30, 2004;
279(5):
3160 - 3168.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. W. Baumann, R. Baur, and E. Sigel
Individual Properties of the Two Functional Agonist Sites in GABAA Receptors
J. Neurosci.,
December 3, 2003;
23(35):
11158 - 11166.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. E. Coyle and D. B. Nikolov
GABARAP: Lessons for Synaptogenesis
Neuroscientist,
June 1, 2003;
9(3):
205 - 216.
[Abstract]
[PDF]
|
 |
|

|
 |

|
 |
 
K. Bollan, D. King, L. A. Robertson, K. Brown, P. M. Taylor, S. J. Moss, and C. N. Connolly
GABAA Receptor Composition Is Determined by Distinct Assembly Signals within alpha and beta Subunits
J. Biol. Chem.,
February 7, 2003;
278(7):
4747 - 4755.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D. E. Raines, R. J. Claycomb, and S. A. Forman
Modulation of GABAA Receptor Function by Nonhalogenated Alkane Anesthetics: The Effects on Agonist Enhancement, Direct Activation, and Inhibition
Anesth. Analg.,
January 1, 2003;
96(1):
112 - 118.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. W. Sawyer, D. C. Chiara, R. W. Olsen, and J. B. Cohen
Identification of the Bovine gamma -Aminobutyric Acid Type A Receptor alpha Subunit Residues Photolabeled by the Imidazobenzodiazepine [3H]Ro15-4513
J. Biol. Chem.,
December 13, 2002;
277(51):
50036 - 50045.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
P. A. Goldstein, F. P. Elsen, S.-W. Ying, C. Ferguson, G. E. Homanics, and N. L. Harrison
Prolongation of Hippocampal Miniature Inhibitory Postsynaptic Currents in Mice Lacking the GABAA Receptor alpha 1 Subunit
J Neurophysiol,
December 1, 2002;
88(6):
3208 - 3217.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. W. Baumann, R. Baur, and E. Sigel
Forced Subunit Assembly in alpha 1beta 2gamma 2 GABAA Receptors. INSIGHT INTO THE ABSOLUTE ARRANGEMENT
J. Biol. Chem.,
November 22, 2002;
277(48):
46020 - 46025.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
E. Palma, V. Esposito, A. M. Mileo, G. Di Gennaro, P. Quarato, F. Giangaspero, C. Scoppetta, P. Onorati, F. Trettel, R. Miledi, et al.
Expression of human epileptic temporal lobe neurotransmitter receptors in Xenopus oocytes: An innovative approach to study epilepsy
PNAS,
November 12, 2002;
99(23):
15078 - 15083.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Scheller and S. A. Forman
Coupled and Uncoupled Gating and Desensitization Effects by Pore Domain Mutations in GABAA Receptors
J. Neurosci.,
October 1, 2002;
22(19):
8411 - 8421.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
I. Sarto, T. Klausberger, N. Ehya, B. Mayer, K. Fuchs, and W. Sieghart
A Novel Site on gamma 3 Subunits Important for Assembly of GABAA Receptors
J. Biol. Chem.,
August 16, 2002;
277(34):
30656 - 30664.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. T. Bianchi, L. Song, H. Zhang, and R. L. Macdonald
Two Different Mechanisms of Disinhibition Produced by GABAA Receptor Mutations Linked to Epilepsy in Humans
J. Neurosci.,
July 1, 2002;
22(13):
5321 - 5327.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. G. Newell and S. M. J. Dunn
Functional Consequences of the Loss of High Affinity Agonist Binding to gamma -Aminobutyric Acid Type A Receptors. IMPLICATIONS FOR RECEPTOR DESENSITIZATION
J. Biol. Chem.,
June 7, 2002;
277(24):
21423 - 21430.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. H. Holden and C. Czajkowski
Different Residues in the GABAA Receptor alpha 1T60-alpha 1K70 Region Mediate GABA and SR-95531 Actions
J. Biol. Chem.,
May 17, 2002;
277(21):
18785 - 18792.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
T. J. Jentsch, V. Stein, F. Weinreich, and A. A. Zdebik
Molecular Structure and Physiological Function of Chloride Channels
Physiol Rev,
April 1, 2002;
82(2):
503 - 568.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S.-A. Thompson, P. B. Wingrove, L. Connelly, P. J. Whiting, and K. A. Wafford
Tracazolate Reveals a Novel Type of Allosteric Interaction with Recombinant gamma -Aminobutyric AcidA Receptors
Mol. Pharmacol.,
April 1, 2002;
61(4):
861 - 869.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. B. Christie, C. P. Miralles, and A. L. De Blas
GABAergic Innervation Organizes Synaptic and Extrasynaptic GABAA Receptor Clustering in Cultured Hippocampal Neurons
J. Neurosci.,
February 1, 2002;
22(3):
684 - 697.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. E. Adkins, G. V. Pillai, J. Kerby, T. P. Bonnert, C. Haldon, R. M. McKernan, J. E. Gonzalez, K. Oades, P. J. Whiting, and P. B. Simpson
alpha 4beta 3delta GABAA Receptors Characterized by Fluorescence Resonance Energy Transfer-derived Measurements of Membrane Potential
J. Biol. Chem.,
October 12, 2001;
276(42):
38934 - 38939.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. Hapfelmeier, R. Haseneder, E. Kochs, M. Beyerle, and W. Zieglgansberger
Coadministered Nitrous Oxide Enhances the Effect of Isoflurane on GABAergic Transmission by an Increase in Open-Channel Block
J. Pharmacol. Exp. Ther.,
July 1, 2001;
298(1):
201 - 208.
[Abstract]
[Full Text]
|
 |
|

|
 |

|
 |
 
A. J. Smith, L. Alder, J. Silk, C. Adkins, A. E. Fletcher, T. Scales, J. Kerby, G. Marshall, K. A. Wafford, R. M. McKernan, et al.
Effect of alpha Subunit on Allosteric Modulation of Ion Channel Function in Stably Expressed Human Recombinant gamma -Aminobutyric AcidA Receptors Determined Using 36Cl Ion Flux
Mol. Pharmacol.,
April 16, 2001;
59(5):
1108 - 1118.
[Abstract]
[Full Text]
|
 |
|

|
 |

|
 |
 
M. I. Strakhova, S. C. Harvey, C. M. Cook, J. M. Cook, and P. Skolnick
A Single Amino Acid Residue on the alpha 5 Subunit (Ile215) Is Essential for Ligand Selectivity at alpha 5beta 3gamma 2gamma -Aminobutyric AcidA Receptors
Mol. Pharmacol.,
April 13, 2001;
58(6):
1434 - 1440.
[Abstract]
[Full Text]
|
 |
|

|
 |

|
 |
 
M. T. Bianchi, K. F. Haas, and R. L. Macdonald
Structural Determinants of Fast Desensitization and Desensitization-Deactivation Coupling in GABAA Receptors
J. Neurosci.,
February 15, 2001;
21(4):
1127 - 1136.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. Hapfelmeier, W. Zieglgansberger, R. Haseneder, H. Schneck, and E. Kochs
Nitrous Oxide and Xenon Increase the Efficacy of GABA at Recombinant Mammalian GABAA Receptors
Anesth. Analg.,
December 1, 2000;
91(6):
1542 - 1549.
[Abstract]
[Full Text]
[PDF]
|
 |
| |