Presented at the Global Medicinal Chemistry and GPCR Summit. To find out more, visit: www.global-engage.com Torsten Schöneberg, from the Rudolf Schönheimer Institute of Biochemistry and University of Leipzig, presents an overview of our current knowledge on aGPCR activation and signal transduction with a focus on the latest findings regarding the interplay between ligand binding, mechanical force, and the tethered agonistic sequence.

1. Targeting Giants
The Pharmacology of Adhesion GPCRs
Torsten Schöneberg
Rudolf Schönheimer Institute
of Biochemistry
Medical Faculty, University of Leipzig
Leipzig, Germany

2. The class of adhesion GPCRs
A
F
lutamate
hodopsin
dhesion
R
G
rizzled/Taste2
ecretinS
lutamateG
R
lutamateG
hodopsinR
lutamateG
A
hodopsinR
lutamateG
dhesionA
hodopsinR
lutamateG
F
dhesionA
hodopsinR
lutamateG
rizzled/Taste2F
dhesionA
hodopsinR
lutamateG
S
rizzled/Taste2F
dhesionA
hodopsinR
lutamateG
ecretinS
F
A
hodopsinR
lutamateG lutamateG
R
lutamateG
hodopsinR
lutamateG
A
R
lutamateG
A
R
lutamateG
F
A
R
(Family C)G
F
A
R
G
S
F
A
R
G
S
F
A
R
G
S ecretin
F
A
R
GG
R
G
R
G
F
R
G
F
R
G
S
F
R
G
A
(Family A)
(Family B)
(Family B)

3. Groups of adhesion GPCRs
Hamann J., et al. Pharm Rev. 2015;67(2):338-67.

4. Giants
- > 750 Mio. years old
- N terminus - up to 5,800 aa
- C terminus - up to 450 aa
- numerous interaction partners
- numerous splice variants
Adhesion GPCR are giants among receptors

5. Evolution of aGPCRs and phenotypes in humans
Signatures of recent selection
- GPR133
- GPR110, GPR111, GPR123
- GPR56, GPR64, GPR126
- EMR1, BAI1, CELSR1 …
Kovacs P, Schöneberg T. Handb Exp Pharmacol. 2016;234:179-217.
Selection in human populations
- GPR133
- GPR110, GPR111, GPR113
- GPR115, GPR64, GPR126
- EMR1, EMR3, BAI3 …
QTL/SNP association
GPR133 - height, weight, lipid, glucose, heart rate, myocardial
infarction
GPR126 - weight, lipid, glucose, insulin, height, pulmonary
function
…

6. GPR133 promotes glioblastoma growth
Bayin et al. Oncogenesis. 2016;5:e263.

7. Adhesion GPCRs couple to G proteins
Bohnekamp J, Schöneberg T. J Biol Chem. 2011;286:41912
SP
P2Y12 N term.
HA tag
FLAG tag
Human GPR133

8. Deletion of the N terminus initiates G-protein coupling
w
t
C
TF
P2Y
12-C
TF
0
200
400
600
800
GPR133
***
***
cAMPaccumulation
[%ofwt]
SP
P2Y12 N term.
HA tag
FLAG tag
SP
HA tag
FLAG tag
CTF P2Y12 CTF

9. Ectodomain as tethered inverse agonist
Hypothetic scenarios of adhesion GPCR activation
Ligand binding
autoprot. cleavage

10. Ectodomain contains a tethered agonist
Hypothetic scenarios of adhesion GPCR activation
Ligand binding
autoprot. cleavage

11. Deletion of the ectodomain activates GPR126
w
t
C
TF
P2Y
12-C
TF
Δ
G
PS-C
TF
P2Y
12-Δ
G
PS-C
TF
0
200
400
600
800
***
***
cAMPaccumulation
[%ofwt]
Liebscher et al. Cell Rep. 2014;9:2018-26.

12. Peptide activation of GPR126

13. Peptide activation of GPR126

14. The ‚Stachel‘ Sequence
EMR1 SSFAVLMAHYDVQ
CD97 SSFAILMAHYDVE
GPR56 TYFAVLMVSSVEV
GPR114 TYFAVLMQLSGDP
GPR115 TSFSILMSSKPVK
GPR116 TSFSILMSPDSPD
GPR126 THFGVLMDLPRSA
GPR133 TNFAILMQVVPLE

15. The ‚Stachel‘ Sequences of aGPCRs
aGPCR Peptide length EC50 reference
GPR126 16 AA > 400 µM Liebscher et al. (2014)
Cell Rep
GPR133 13 AA > 400 µM Liebscher et al. (2014)
Cell Rep
GPR56 7 AA 35 µM Stoveken et al. (2015)
PNAS
GPR110 10 AA 37 µM Stoveken et al. (2015)
PNAS
GPR64 15 AA 77 µM Demberg et al. (2015)
BBRC
GPR114 18 AA 100 µM Wilde et al. (2015)
Faseb J
Lat-1 12 AA Not determined Müller et al. (2015)
PLoS Genet

16. ‚Stachel‘ sequence in an aGPCR model

17. Mogha et al. J Neurosci. 2013; 33:17976-85.
GPR126 is a Gs protein-coupled receptor

18. Liebscher et al. Cell Rep. 2014;9:2018-26.
Rescue of myelination phenotype in GPR126-mutant zebrafish
Stachel sequence

19. Stachel peptide specificity
Hamann J., et al. Pharm Rev. 2015;67(2):338-67.

20. Stachel-derived peptide specificity
Demberg et al. J Biol Chem. 2017 292:4383-94
0
5
10
15
20
25
30
pGPR64 - + - - - + - - - + - - - + - -
pGPR126 - - + - - - + - - - + - - - + -
pGPR110 - - - + - - - + - - - + - - - +
pcDps GPR64 GPR126 GPR110
***
***
**
**
*
***
cAMPaccumulation
(x-foldoveremptyvector,
nopeptide)

21. Tethered agonism in GPCRs
protease
GPHs
thyreostimulin
matrix proteins
cell-cell contacts
proteases
mechanical force ?
PARs GPHRs Adhesion GPCRs
p10 StachelTRAP
relaxin
RXFP1
LDLa

22. GPR126 is activated by shaking
Petersen et al. Neuron 2015;85:755-69.

23. Small Ligands of aGPCRs
Dihydromunduletone Stoveken et al. Mol Pharmacol. 2016;90:214-24.
Antagonist at GPR56 and GPR114
Gupte et al. FEBS Lett. 2012;586:1214-9.

24. cis/trans signaling of adhesion GPCRs
trans signaling
(other partners)
cis signaling
(G proteins)
(and others, e.g. ELMO/Dock)

25. University of Leipzig, Germany
Ines Liebscher
Julia Schön
Lilian Demberg
Caro Wilde
Nina Auerbach
Vera Lede
Christin Schröck
Simone Prömel
Jana Winkler
Antje Müller
Charite Berlin, Germany
Gunnar Kleinau
Vollum Institute
Portland, OR, USA
Kelly Monk
Acknowledgment
http://www.adhesiongpcr.org

26. Presented at the Global Medicinal
Chemistry and GPCR Summit.
To find out more, visit:
www.global-engage.com