Receptor activity-modifying proteins 2 and 3 generate adrenomedullin receptor subtypes with distinct molecular properties

Watkins, Harriet A., Chakravarthy, Madhuri, Abhayawardana, Rekhati S., Gingell, Joseph J., Garelja, Michael, Pardamwar, Meenakshi, McElhinney, James M.W.R., Lathbridge, Alex, Constantine, Arran, Harris, Paul W.R., Yuen, Tsz-Ying, Brimble, Margaret A., Barwell, James, Poyner, David R., Woolley, Michael J., Conner, Alex C., Pioszak, Augen A., Reynolds, Christopher A. and Hay, Debbie L. (2016). Receptor activity-modifying proteins 2 and 3 generate adrenomedullin receptor subtypes with distinct molecular properties. Journal of Biological Chemistry, 291 (22), pp. 11657-11675.

Abstract

Adrenomedullin (AM) is a peptide hormone with numerous effects in the vascular systems. AM signals through the AM1 and AM2 receptors formed by the obligate heterodimerization of a G protein-coupled receptor, the calcitonin receptor-like receptor (CLR), and receptor activity-modifying proteins (RAMP) 2 and 3, respectively. These different CLR-RAMP interactions yield discrete receptor pharmacology and physiological effects. The effective design of therapeutics that target the individual AM receptors is dependent on understanding the molecular details of the effects of RAMPs on CLR. To understand the role of RAMPs 2 and 3 on the activation and conformation of the CLR subunit of AM receptors we mutated 68 individual amino acids in the juxtamembrane region of CLR, a key region for activation of AM receptors and determined the effects on cAMP signalling. Sixteen CLR mutations had differential effects between the AM1 and AM2 receptors. Accompanying this, independent molecular modelling of the full-length AM-bound AM1 and AM2 receptors predicted differences in the binding pocket, and differences in the electrostatic potential of the two AM receptors. Druggability analysis indicated unique features that could be used to develop selective small molecule ligands for each receptor. The interaction of RAMP2 or RAMP3 with CLR induces conformational variation in the juxtamembrane region, yielding distinct binding pockets, probably via an allosteric mechanism. These subtype-specific differences have implications for the design of therapeutics aimed at specific AM receptors and for understanding the mechanisms by which accessory proteins affect G protein-coupled receptor function.

Publication DOI: https://doi.org/10.1074/jbc.M115.688218
Divisions: Life & Health Sciences > Pharmacy
Life & Health Sciences > Biosciences
Life & Health Sciences
Additional Information: Final version free via Creative Commons CC-BY license. Funding: BBSRC (BB/M006883/1; BB/M007529/1 and BB/M000176/1) Supplemental material: http://www.jbc.org/content/suppl/2016/04/13/M115.688218.DC1.htm
Uncontrolled Keywords: allosteric regulation,cardiovascular disease,conformational change,G protein-coupled receptor,GPCR,molecular modeling,RAMP,adrenomedullin,extracellular loops,receptor activity-modifying protein,Biochemistry,Cell Biology,Molecular Biology
Full Text Link: http://www.jbc. ... jbc.M115.688218
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Date: 2016-05-27
Authors: Watkins, Harriet A.
Chakravarthy, Madhuri
Abhayawardana, Rekhati S.
Gingell, Joseph J.
Garelja, Michael
Pardamwar, Meenakshi
McElhinney, James M.W.R.
Lathbridge, Alex
Constantine, Arran
Harris, Paul W.R.
Yuen, Tsz-Ying
Brimble, Margaret A.
Barwell, James
Poyner, David R. ( 0000-0003-1590-112X)
Woolley, Michael J.
Conner, Alex C.
Pioszak, Augen A.
Reynolds, Christopher A.
Hay, Debbie L.

Download

[img]

Version: Published Version

License: Creative Commons Attribution


[img]

Access Restriction: Restricted to Repository staff only

License: Creative Commons Attribution


Export / Share Citation


Statistics

Additional statistics for this record