Abstract
The activation of G proteins by G protein-coupled receptors (GPCRs) underlies the majority of transmembrane signaling by hormones and neurotransmitters. Recent structures of GPCR-G protein complexes obtained by crystallography and cryoelectron microscopy (cryo-EM) reveal similar interactions between GPCRs and the alpha subunit of different G protein isoforms. While some G protein subtype-specific differences are observed, there is no clear structural explanation for G protein subtype-selectivity. All of these complexes are stabilized in the nucleotide-free state, a condition that does not exist in living cells. In an effort to better understand the structural basis of coupling specificity, we used time-resolved structural mass spectrometry techniques to investigate GPCR-G protein complex formation and G-protein activation. Our results suggest that coupling specificity is determined by one or more transient intermediate states that serve as selectivity filters and precede the formation of the stable nucleotide-free GPCR-G protein complexes observed in crystal and cryo-EM structures. A time-resolved look at how a GPCR engages a G protein reveals intermediates in the process that dictate both specificity for the interaction and the initial steps kicking off downstream signaling.
| Original language | English |
|---|---|
| Pages (from-to) | 1232-1242 |
| Number of pages | 11 |
| Journal | Cell |
| Volume | 177 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 16 May 2019 |
| Externally published | Yes |
Keywords
- conformation
- dynamics
- G protein
- G protein-coupled receptor
- hydrogen/deuterium exchange mass spectrometry
- hydroxyl radical footprinting mass spectrometry
Cite this
}
Assembly of a GPCR-G Protein Complex. / Du, Yang; Duc, Nguyen Minh; Rasmussen, Søren G.F.; Hilger, Daniel; Kubiak, Xavier; Wang, Liwen; Bohon, Jennifer; Kim, Hee Ryung; Wegrecki, Marcin; Asuru, Awuri; Jeong, Kyung Min; Lee, Jeongmi; Chance, Mark R.; Lodowski, David T.; Chung, Ka Young.
In: Cell, Vol. 177, No. 5, 16.05.2019, p. 1232-1242.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Assembly of a GPCR-G Protein Complex
AU - Du, Yang
AU - Duc, Nguyen Minh
AU - Rasmussen, Søren G.F.
AU - Hilger, Daniel
AU - Kubiak, Xavier
AU - Wang, Liwen
AU - Bohon, Jennifer
AU - Kim, Hee Ryung
AU - Wegrecki, Marcin
AU - Asuru, Awuri
AU - Jeong, Kyung Min
AU - Lee, Jeongmi
AU - Chance, Mark R.
AU - Lodowski, David T.
AU - Chung, Ka Young
PY - 2019/5/16
Y1 - 2019/5/16
N2 - The activation of G proteins by G protein-coupled receptors (GPCRs) underlies the majority of transmembrane signaling by hormones and neurotransmitters. Recent structures of GPCR-G protein complexes obtained by crystallography and cryoelectron microscopy (cryo-EM) reveal similar interactions between GPCRs and the alpha subunit of different G protein isoforms. While some G protein subtype-specific differences are observed, there is no clear structural explanation for G protein subtype-selectivity. All of these complexes are stabilized in the nucleotide-free state, a condition that does not exist in living cells. In an effort to better understand the structural basis of coupling specificity, we used time-resolved structural mass spectrometry techniques to investigate GPCR-G protein complex formation and G-protein activation. Our results suggest that coupling specificity is determined by one or more transient intermediate states that serve as selectivity filters and precede the formation of the stable nucleotide-free GPCR-G protein complexes observed in crystal and cryo-EM structures. A time-resolved look at how a GPCR engages a G protein reveals intermediates in the process that dictate both specificity for the interaction and the initial steps kicking off downstream signaling.
AB - The activation of G proteins by G protein-coupled receptors (GPCRs) underlies the majority of transmembrane signaling by hormones and neurotransmitters. Recent structures of GPCR-G protein complexes obtained by crystallography and cryoelectron microscopy (cryo-EM) reveal similar interactions between GPCRs and the alpha subunit of different G protein isoforms. While some G protein subtype-specific differences are observed, there is no clear structural explanation for G protein subtype-selectivity. All of these complexes are stabilized in the nucleotide-free state, a condition that does not exist in living cells. In an effort to better understand the structural basis of coupling specificity, we used time-resolved structural mass spectrometry techniques to investigate GPCR-G protein complex formation and G-protein activation. Our results suggest that coupling specificity is determined by one or more transient intermediate states that serve as selectivity filters and precede the formation of the stable nucleotide-free GPCR-G protein complexes observed in crystal and cryo-EM structures. A time-resolved look at how a GPCR engages a G protein reveals intermediates in the process that dictate both specificity for the interaction and the initial steps kicking off downstream signaling.
KW - conformation
KW - dynamics
KW - G protein
KW - G protein-coupled receptor
KW - hydrogen/deuterium exchange mass spectrometry
KW - hydroxyl radical footprinting mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=85065516910&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2019.04.022
DO - 10.1016/j.cell.2019.04.022
M3 - Article
AN - SCOPUS:85065516910
VL - 177
SP - 1232
EP - 1242
JO - Cell
JF - Cell
SN - 0092-8674
IS - 5
ER -