Abstract
Seven transmembrane (7TM) or G protein-coupled receptors constitute a large superfamily of cell surface receptors sharing a structural motif of seven transmembrane spanning alpha helices. Their activation mechanism most likely involves concerted movements of the transmembrane helices, but remains to be completely resolved. Evolutionary Trace (ET) analysis is a computational method, which identifies clusters of functionally important residues by integrating information on evolutionary important residue variations with receptor structure. Combined with known mutational data, ET predicted a patch of residues in the cytoplasmic parts of TM2, TM3, and TM6 to form an activation switch that is common to all family A 7TM receptors. We tested this hypothesis in the rat Angiotensin II (Ang II) type 1a (AT1a) receptor. The receptor has important roles in the cardiovascular system, but has also frequently been applied as a model for 7TM receptor activation and signaling. Six mutations: F66A, L67R, L70R, L119R, D125A, and I245F were targeted to the putative switch and assayed for changes in activation state by their ligand binding, signaling, and trafficking properties. All but one receptor mutant (that was not expressed well) displayed phenotypes associated with changed activation state, such as increased agonist affinity or basal activity, promiscuous activation, or constitutive internalization highlighting the importance of testing different signaling pathways. We conclude that this evolutionary important patch mediates interactions important for maintaining the inactive state. More broadly, these observations in the AT1 receptor are consistent with computational predictions of a generic role for this patch in 7TM receptor activation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 86-94 |
| Number of pages | 9 |
| Journal | Biochemical Pharmacology |
| Volume | 80 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jul 1 2010 |
| Externally published | Yes |
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Keywords
- 7TM receptor
- Angiotensin
- Constitutive activity
- Evolutionary Trace
- GPCR
ASJC Scopus subject areas
- Biochemistry
- Pharmacology
Cite this
An Angiotensin II type 1 receptor activation switch patch revealed through Evolutionary Trace analysis. / Bonde, Marie Mi; Yao, Rong; Ma, Jian Nong; Madabushi, Srinivasan; Haunsø, Stig; Burstein, Ethan S.; Whistler, Jennifer; Sheikh, Søren P.; Lichtarge, Olivier; Hansen, Jakob Lerche.
In: Biochemical Pharmacology, Vol. 80, No. 1, 01.07.2010, p. 86-94.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - An Angiotensin II type 1 receptor activation switch patch revealed through Evolutionary Trace analysis
AU - Bonde, Marie Mi
AU - Yao, Rong
AU - Ma, Jian Nong
AU - Madabushi, Srinivasan
AU - Haunsø, Stig
AU - Burstein, Ethan S.
AU - Whistler, Jennifer
AU - Sheikh, Søren P.
AU - Lichtarge, Olivier
AU - Hansen, Jakob Lerche
PY - 2010/7/1
Y1 - 2010/7/1
N2 - Seven transmembrane (7TM) or G protein-coupled receptors constitute a large superfamily of cell surface receptors sharing a structural motif of seven transmembrane spanning alpha helices. Their activation mechanism most likely involves concerted movements of the transmembrane helices, but remains to be completely resolved. Evolutionary Trace (ET) analysis is a computational method, which identifies clusters of functionally important residues by integrating information on evolutionary important residue variations with receptor structure. Combined with known mutational data, ET predicted a patch of residues in the cytoplasmic parts of TM2, TM3, and TM6 to form an activation switch that is common to all family A 7TM receptors. We tested this hypothesis in the rat Angiotensin II (Ang II) type 1a (AT1a) receptor. The receptor has important roles in the cardiovascular system, but has also frequently been applied as a model for 7TM receptor activation and signaling. Six mutations: F66A, L67R, L70R, L119R, D125A, and I245F were targeted to the putative switch and assayed for changes in activation state by their ligand binding, signaling, and trafficking properties. All but one receptor mutant (that was not expressed well) displayed phenotypes associated with changed activation state, such as increased agonist affinity or basal activity, promiscuous activation, or constitutive internalization highlighting the importance of testing different signaling pathways. We conclude that this evolutionary important patch mediates interactions important for maintaining the inactive state. More broadly, these observations in the AT1 receptor are consistent with computational predictions of a generic role for this patch in 7TM receptor activation.
AB - Seven transmembrane (7TM) or G protein-coupled receptors constitute a large superfamily of cell surface receptors sharing a structural motif of seven transmembrane spanning alpha helices. Their activation mechanism most likely involves concerted movements of the transmembrane helices, but remains to be completely resolved. Evolutionary Trace (ET) analysis is a computational method, which identifies clusters of functionally important residues by integrating information on evolutionary important residue variations with receptor structure. Combined with known mutational data, ET predicted a patch of residues in the cytoplasmic parts of TM2, TM3, and TM6 to form an activation switch that is common to all family A 7TM receptors. We tested this hypothesis in the rat Angiotensin II (Ang II) type 1a (AT1a) receptor. The receptor has important roles in the cardiovascular system, but has also frequently been applied as a model for 7TM receptor activation and signaling. Six mutations: F66A, L67R, L70R, L119R, D125A, and I245F were targeted to the putative switch and assayed for changes in activation state by their ligand binding, signaling, and trafficking properties. All but one receptor mutant (that was not expressed well) displayed phenotypes associated with changed activation state, such as increased agonist affinity or basal activity, promiscuous activation, or constitutive internalization highlighting the importance of testing different signaling pathways. We conclude that this evolutionary important patch mediates interactions important for maintaining the inactive state. More broadly, these observations in the AT1 receptor are consistent with computational predictions of a generic role for this patch in 7TM receptor activation.
KW - 7TM receptor
KW - Angiotensin
KW - Constitutive activity
KW - Evolutionary Trace
KW - GPCR
UR - http://www.scopus.com/inward/record.url?scp=77952290328&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77952290328&partnerID=8YFLogxK
U2 - 10.1016/j.bcp.2010.03.006
DO - 10.1016/j.bcp.2010.03.006
M3 - Article
C2 - 20227396
AN - SCOPUS:77952290328
VL - 80
SP - 86
EP - 94
JO - Biochemical Pharmacology
JF - Biochemical Pharmacology
SN - 0006-2952
IS - 1
ER -