Metabotropic GABAB receptors mediate inhibitory neurotransmission in the mammalian brain and are central to its function. GABAB receptors are G-protein coupled receptors (GPCRs) that regulate the activity of Ca2+ and K+ channels, and inhibit the function of adenylyl cyclase. Malfunction of GABAB receptors has been implicated in the development of a number of diseases including spasticity, epilepsy and pain. Therefore, elucidating the structure and function of GABAB receptors would assist the design of valuable therapeutic agents. Unlike most GPCRs, GABAB receptors function as a heterodimeric assembly of GBR1 and GBR2 subunits. Heterodimerization is required for the trans-activation of these receptors, where GBR1 is responsible for ligand-binding and GBR2 is involved in G-protein coupling. The first part of this proposal is aimed at understanding the role of GBR1 and GBR2 ectodomains in the activation process. We propose to solve the structures of the individual subunits and study their interactions using biochemical methods.
The second aim i s to reveal the structural basis of ligand-binding and dimerization of an ectodomain heterodimer, and determine the affinity and specificity of receptor-ligand interactions.
Our third aim i s to understand the molecular basis of heterodimerization by the intracellular coiled-coil domains and their role in receptor assembly and trafficking. The last part of the proposed research is focused on understanding transmembrane signal transduction mechanism using full-length receptors. We propose to carry out functional studies of wild-type and mutant receptors using whole cell patch clamp methodology. Mutations that affect ligand binding, receptor heterodimerization, and receptor activation will be designed based on the available structural information and homology models. A combination of structural and functional analysis of GABAB receptors will advance our understanding of the molecular basis of GABA action and the activation mechanisms particular to GPCR heterodimers.

Public Health Relevance

Malfunction of metabotropic GABA receptors leads to neurological disorders in human. We are pursuing structural models for different components of GABA receptors in order to understand their roles in receptor function. The resulting atomic structures may assist the design of novel therapeutic agents.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM088454-05
Application #
8513349
Study Section
Special Emphasis Panel (ZRG1-MDCN-C (02))
Program Officer
Ainsztein, Alexandra M
Project Start
2009-08-15
Project End
2014-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
5
Fiscal Year
2013
Total Cost
$287,797
Indirect Cost
$109,041
Name
Columbia University (N.Y.)
Department
Pharmacology
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Burmakina, Svetlana; Geng, Yong; Chen, Yan et al. (2014) Heterodimeric coiled-coil interactions of human GABAB receptor. Proc Natl Acad Sci U S A 111:6958-63
Geng, Yong; Bush, Martin; Mosyak, Lidia et al. (2013) Structural mechanism of ligand activation in human GABA(B) receptor. Nature 504:254-9