TY - JOUR
T1 - Rational Design of Nanobody80 Loop Peptidomimetics
T2 - Towards Biased β2 Adrenergic Receptor Ligands
AU - Martin, Charlotte
AU - Moors, Samuel L C
AU - Danielsen, Mia
AU - Betti, Cecilia
AU - Fabris, Cecilia
AU - Sejer Pedersen, Daniel
AU - Pardon, Els
AU - Peyressatre, Marion
AU - Fehér, Krisztina
AU - Martins, José C
AU - Mosolff Mathiesen, Jesper
AU - Morris, May C
AU - Devoogdt, Nick
AU - Caveliers, Vicky
AU - De Proft, Frank
AU - Steyaert, Jan
AU - Ballet, Steven
N1 - © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2017/7/18
Y1 - 2017/7/18
N2 - G protein-coupled receptors (GPCRs) play an important role in many cellular responses; as such, their mechanism of action is of utmost interest. To gain insight into the active conformation of GPCRs, the X-ray crystal structures of nanobody (Nb)-stabilized β2 -adrenergic receptor (β2 AR) have been reported. Nb80, in particular, is able to bind the intracellular G protein binding site of β2 AR and stabilize the receptor in an active conformation. Within Nb80, the complementarity-determining region 3 (CDR3) is responsible for most of the binding interactions. Hence, we hypothesized that peptidomimetics of the CDR3 loop might be sufficient for binding to the receptor, inhibiting the interaction of β2 AR with intracellular GPCR interacting proteins (e.g., G proteins). Based on previous crystallographic data, a set of peptidomimetics were synthesized that, similar to the Nb80 CDR3 loop, adopt a β-hairpin conformation. Syntheses, conformational analysis, binding and functional in vitro assays, as well as internalization experiments, were performed. We demonstrate that peptidomimetics can structurally mimic the CDR3 loop of a nanobody and its function by inhibiting G protein coupling as measured by partial inhibition of cAMP production.
AB - G protein-coupled receptors (GPCRs) play an important role in many cellular responses; as such, their mechanism of action is of utmost interest. To gain insight into the active conformation of GPCRs, the X-ray crystal structures of nanobody (Nb)-stabilized β2 -adrenergic receptor (β2 AR) have been reported. Nb80, in particular, is able to bind the intracellular G protein binding site of β2 AR and stabilize the receptor in an active conformation. Within Nb80, the complementarity-determining region 3 (CDR3) is responsible for most of the binding interactions. Hence, we hypothesized that peptidomimetics of the CDR3 loop might be sufficient for binding to the receptor, inhibiting the interaction of β2 AR with intracellular GPCR interacting proteins (e.g., G proteins). Based on previous crystallographic data, a set of peptidomimetics were synthesized that, similar to the Nb80 CDR3 loop, adopt a β-hairpin conformation. Syntheses, conformational analysis, binding and functional in vitro assays, as well as internalization experiments, were performed. We demonstrate that peptidomimetics can structurally mimic the CDR3 loop of a nanobody and its function by inhibiting G protein coupling as measured by partial inhibition of cAMP production.
KW - Journal Article
U2 - 10.1002/chem.201701321
DO - 10.1002/chem.201701321
M3 - Journal article
C2 - 28449310
SN - 0947-6539
VL - 23
SP - 9632
EP - 9640
JO - Chemistry: A European Journal
JF - Chemistry: A European Journal
IS - 40
ER -