Structural and functional evidence for testosterone activation of GPRC6A in peripheral tissues

Min Pi, Karan Kapoor, Yunpeng Wu, Ruisong Ye, Susan E. Senogles, Satoru K. Nishimoto, Dong Jin Hwang, Duane D. Miller, Ramesh Narayanan, Jeremy C. Smith, Jerome Baudry, L. D.Arryl Quarles

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

G protein-coupled receptor (GPCR) family C group 6 member A (GPRC6A) is a multiligand GPCR that is activated by cations, L-amino acids, and osteocalcin. GPRC6A plays an important role in the regulation of testosterone (T) production and energy metabolism in mice. T has rapid, transcription-independent (nongenomic) effects that are mediated by a putative GPCR. We previously found that T can activate GPRC6A in vitro, but the possibility that T is a ligand for GPRC6A remains controversial. Here, we demonstrate direct T binding to GPRC6A and construct computational structural models of GPRC6Athat are used to identify potential binding poses of T. Mutations of the predicted binding site residues were experimentally found to block T activation of GPRC6A, in agreement with the modeling. Using Gpr6ca-/- mice, we confirmed that loss of GPRC6A resulted in loss of T rapid signaling responses and elucidated several biological functions regulated by GPRC6A-dependent T rapid signaling, including T stimulation of insulin secretion in pancreatic islets and enzyme expression involved in the biosynthesis of T in Leydig cells. Finally, we identified a stereo-specific effect of an R-isomer of a selective androgen receptor modulatorthat is predicted to bind to and shown to activate GPRC6A but not androgen receptor. Together, our data show that GPRC6A directly mediates the rapid signaling response to T and uncovers previously unrecognized endocrine networks.

Original languageEnglish (US)
Pages (from-to)1759-1773
Number of pages15
JournalMolecular Endocrinology
Volume29
Issue number12
DOIs
StatePublished - Dec 2015

Fingerprint

G-Protein-Coupled Receptors
Testosterone
Androgen Receptors
Leydig Cells
Structural Models
Osteocalcin
Islets of Langerhans
Energy Metabolism
Cations
Binding Sites
Insulin
Ligands
Amino Acids
Mutation
Enzymes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Structural and functional evidence for testosterone activation of GPRC6A in peripheral tissues. / Pi, Min; Kapoor, Karan; Wu, Yunpeng; Ye, Ruisong; Senogles, Susan E.; Nishimoto, Satoru K.; Hwang, Dong Jin; Miller, Duane D.; Narayanan, Ramesh; Smith, Jeremy C.; Baudry, Jerome; Quarles, L. D.Arryl.

In: Molecular Endocrinology, Vol. 29, No. 12, 12.2015, p. 1759-1773.

Research output: Contribution to journalArticle

@article{9a8de0eb8fa9479992a4d1ed9b65b60b,
title = "Structural and functional evidence for testosterone activation of GPRC6A in peripheral tissues",
abstract = "G protein-coupled receptor (GPCR) family C group 6 member A (GPRC6A) is a multiligand GPCR that is activated by cations, L-amino acids, and osteocalcin. GPRC6A plays an important role in the regulation of testosterone (T) production and energy metabolism in mice. T has rapid, transcription-independent (nongenomic) effects that are mediated by a putative GPCR. We previously found that T can activate GPRC6A in vitro, but the possibility that T is a ligand for GPRC6A remains controversial. Here, we demonstrate direct T binding to GPRC6A and construct computational structural models of GPRC6Athat are used to identify potential binding poses of T. Mutations of the predicted binding site residues were experimentally found to block T activation of GPRC6A, in agreement with the modeling. Using Gpr6ca-/- mice, we confirmed that loss of GPRC6A resulted in loss of T rapid signaling responses and elucidated several biological functions regulated by GPRC6A-dependent T rapid signaling, including T stimulation of insulin secretion in pancreatic islets and enzyme expression involved in the biosynthesis of T in Leydig cells. Finally, we identified a stereo-specific effect of an R-isomer of a selective androgen receptor modulatorthat is predicted to bind to and shown to activate GPRC6A but not androgen receptor. Together, our data show that GPRC6A directly mediates the rapid signaling response to T and uncovers previously unrecognized endocrine networks.",
author = "Min Pi and Karan Kapoor and Yunpeng Wu and Ruisong Ye and Senogles, {Susan E.} and Nishimoto, {Satoru K.} and Hwang, {Dong Jin} and Miller, {Duane D.} and Ramesh Narayanan and Smith, {Jeremy C.} and Jerome Baudry and Quarles, {L. D.Arryl}",
year = "2015",
month = "12",
doi = "10.1210/me.2015-1161",
language = "English (US)",
volume = "29",
pages = "1759--1773",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "12",

}

TY - JOUR

T1 - Structural and functional evidence for testosterone activation of GPRC6A in peripheral tissues

AU - Pi, Min

AU - Kapoor, Karan

AU - Wu, Yunpeng

AU - Ye, Ruisong

AU - Senogles, Susan E.

AU - Nishimoto, Satoru K.

AU - Hwang, Dong Jin

AU - Miller, Duane D.

AU - Narayanan, Ramesh

AU - Smith, Jeremy C.

AU - Baudry, Jerome

AU - Quarles, L. D.Arryl

PY - 2015/12

Y1 - 2015/12

N2 - G protein-coupled receptor (GPCR) family C group 6 member A (GPRC6A) is a multiligand GPCR that is activated by cations, L-amino acids, and osteocalcin. GPRC6A plays an important role in the regulation of testosterone (T) production and energy metabolism in mice. T has rapid, transcription-independent (nongenomic) effects that are mediated by a putative GPCR. We previously found that T can activate GPRC6A in vitro, but the possibility that T is a ligand for GPRC6A remains controversial. Here, we demonstrate direct T binding to GPRC6A and construct computational structural models of GPRC6Athat are used to identify potential binding poses of T. Mutations of the predicted binding site residues were experimentally found to block T activation of GPRC6A, in agreement with the modeling. Using Gpr6ca-/- mice, we confirmed that loss of GPRC6A resulted in loss of T rapid signaling responses and elucidated several biological functions regulated by GPRC6A-dependent T rapid signaling, including T stimulation of insulin secretion in pancreatic islets and enzyme expression involved in the biosynthesis of T in Leydig cells. Finally, we identified a stereo-specific effect of an R-isomer of a selective androgen receptor modulatorthat is predicted to bind to and shown to activate GPRC6A but not androgen receptor. Together, our data show that GPRC6A directly mediates the rapid signaling response to T and uncovers previously unrecognized endocrine networks.

AB - G protein-coupled receptor (GPCR) family C group 6 member A (GPRC6A) is a multiligand GPCR that is activated by cations, L-amino acids, and osteocalcin. GPRC6A plays an important role in the regulation of testosterone (T) production and energy metabolism in mice. T has rapid, transcription-independent (nongenomic) effects that are mediated by a putative GPCR. We previously found that T can activate GPRC6A in vitro, but the possibility that T is a ligand for GPRC6A remains controversial. Here, we demonstrate direct T binding to GPRC6A and construct computational structural models of GPRC6Athat are used to identify potential binding poses of T. Mutations of the predicted binding site residues were experimentally found to block T activation of GPRC6A, in agreement with the modeling. Using Gpr6ca-/- mice, we confirmed that loss of GPRC6A resulted in loss of T rapid signaling responses and elucidated several biological functions regulated by GPRC6A-dependent T rapid signaling, including T stimulation of insulin secretion in pancreatic islets and enzyme expression involved in the biosynthesis of T in Leydig cells. Finally, we identified a stereo-specific effect of an R-isomer of a selective androgen receptor modulatorthat is predicted to bind to and shown to activate GPRC6A but not androgen receptor. Together, our data show that GPRC6A directly mediates the rapid signaling response to T and uncovers previously unrecognized endocrine networks.

UR - http://www.scopus.com/inward/record.url?scp=84948973471&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84948973471&partnerID=8YFLogxK

U2 - 10.1210/me.2015-1161

DO - 10.1210/me.2015-1161

M3 - Article

C2 - 26440882

AN - SCOPUS:84948973471

VL - 29

SP - 1759

EP - 1773

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 12

ER -