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@ARTICLE{Watkins:139938,
author = {Watkins, HA and Au, M and Bobby, R and Archbold, JK and
Abdul-Manan, N and Moore, JM and Middleditch, MJ and
Williams, GM and Brimble, MA and Dingley, Andrew and Hay,
DL},
title = {{I}dentification of key residues involved in adrenomedullin
binding to the {AM} 1 receptor},
journal = {British journal of pharmacology},
volume = {169},
number = {1},
issn = {0007-1188},
address = {Malden, MA},
publisher = {Wiley},
reportid = {FZJ-2013-05905},
pages = {143 - 155},
year = {2013},
abstract = {BACKGROUND AND PURPOSE:Adrenomedullin (AM) is a peptide
hormone whose receptors are members of the class B GPCR
family. They comprise a heteromer between the GPCR, the
calcitonin receptor-like receptor and one of the receptor
activity-modifying proteins 1-3. AM plays a significant role
in angiogenesis and its antagonist fragment AM22-52 can
inhibit blood vessel and tumour growth. The mechanism by
which AM interacts with its receptors is
unknown.EXPERIMENTAL APPROACH:We determined the AM22-52
binding epitope for the AM1 receptor extracellular domain
using biophysical techniques, heteronuclear magnetic
resonance spectroscopy and alanine scanning.KEY
RESULTS:Chemical shift perturbation experiments located the
main binding epitope for AM22-52 at the AM1 receptor to the
C-terminal 8 amino acids. Isothermal titration calorimetry
of AM22-52 alanine-substituted peptides indicated that Y52,
G51 and I47 are essential for AM1 receptor binding and that
K46 and P49 and R44 have a smaller role to play.
Characterization of these peptides at the full-length AM
receptors was assessed in Cos7 cells by cAMP assay. This
confirmed the essential role of Y52, G51 and I47 in binding
to the AM1 receptor, with their substitution resulting in
≥100-fold reduction in antagonist potency compared with
AM22-52 . R44A, K46A, S48A and P49A AM22-52 decreased
antagonist potency by approximately 10-fold.CONCLUSIONS AND
IMPLICATIONS:This study localizes the main binding epitope
of AM22-52 to its C-terminal amino acids and distinguishes
essential residues involved in this binding. This will
inform the development of improved AM receptor antagonists.},
cin = {ICS-6},
ddc = {610},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {452 - Structural Biology (POF2-452)},
pid = {G:(DE-HGF)POF2-452},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000317679000011},
doi = {10.1111/bph.12118},
url = {https://juser.fz-juelich.de/record/139938},
}