Characterisation of the adiponectin receptors: Cell-surface expression and signal transduction of AdipoR1 and AdipoR2 are palmitoylation-dependent

Sahar Keshvari, Darren Henstridge, Mark Adams, Hayley M. O'Neill, John Hooper, Mark A. Febbraio, Jonathan P Whitehead

Research output: Contribution to journalMeeting AbstractResearchpeer-review

Abstract

The adiponectin axis is a major regulator of metabolic, cardiovascular and inflammatory tone. Current understanding of the adiponectin receptors, R1 and R2, is rudimentary constraining our ability to target these atypical seven transmembrane receptors. Hence, we have started to characterise R1 and R2.We previously reported that R1 exhibits robust (60%) cell-surface expression (CSE) under steady-state conditions (no serum starvation) whilst R2 is undetectable and showed these differences were predicated by the non-conserved N-terminal, intracellular trunks [1]. Here we hypothesised that (i) CSE of R1/R2 is regulated, (ii) R1/R2 are subject to palmitoylation and this regulates CSE/function (as in GPCRs). To address these hypotheses we have employed bioinformatics, cell-based and in vivo approaches. We demonstrate that (i) serum starvation increases R2 CSE and adiponectin stimulates internalisation of R1/R2 (all p < 0.05). (ii) Bioinformatics analysis revealed a putative conserved ‘canonical’ palmitoylation site in the juxtamembrane region of R1(124) and R2(135), plus additional non-conserved sites. This was confirmed using mutagenesis/acylbiotinyl exchange chemistry. Palmitoylation of R1(124) or R2(135) was shown to be required for efficient CSE of R1 and R2 and coupling to downstream signalling effectors including AMPK, AKT and ERK in cell lines and in vivo in mouse skeletal muscle respectively (all p < 0.05). These results provide increasing evidence that CSE of R1 and R2 is under complex, differential regulation and is essential for coupling adiponectin to downstream signalling effectors. Further studies are required to elaborate the detailed molecular mechanisms that may provide novel therapeutic opportunities.
Original languageEnglish
Pages (from-to)114-115
JournalObesity Research and Clinical Practice
Volume8
Issue numberSuppl 1
DOIs
Publication statusPublished - Dec 2014
EventAnnual Scientific Meeting of the Australian and New Zealand Obesity Society in conjunction with the Australian Lifestyle Medicine Association - Novotel on Brighton Beach, Sydney, Australia
Duration: 16 Oct 201418 Oct 2014

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Adiponectin Receptors
Lipoylation
Signal Transduction
Adiponectin
AMP-Activated Protein Kinases
Starvation
Computational Biology
Serum
Mutagenesis
Cell Line

Cite this

Keshvari, Sahar ; Henstridge, Darren ; Adams, Mark ; O'Neill, Hayley M. ; Hooper, John ; Febbraio, Mark A. ; Whitehead, Jonathan P. / Characterisation of the adiponectin receptors: Cell-surface expression and signal transduction of AdipoR1 and AdipoR2 are palmitoylation-dependent. In: Obesity Research and Clinical Practice. 2014 ; Vol. 8, No. Suppl 1. pp. 114-115.
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title = "Characterisation of the adiponectin receptors: Cell-surface expression and signal transduction of AdipoR1 and AdipoR2 are palmitoylation-dependent",
abstract = "The adiponectin axis is a major regulator of metabolic, cardiovascular and inflammatory tone. Current understanding of the adiponectin receptors, R1 and R2, is rudimentary constraining our ability to target these atypical seven transmembrane receptors. Hence, we have started to characterise R1 and R2.We previously reported that R1 exhibits robust (60{\%}) cell-surface expression (CSE) under steady-state conditions (no serum starvation) whilst R2 is undetectable and showed these differences were predicated by the non-conserved N-terminal, intracellular trunks [1]. Here we hypothesised that (i) CSE of R1/R2 is regulated, (ii) R1/R2 are subject to palmitoylation and this regulates CSE/function (as in GPCRs). To address these hypotheses we have employed bioinformatics, cell-based and in vivo approaches. We demonstrate that (i) serum starvation increases R2 CSE and adiponectin stimulates internalisation of R1/R2 (all p < 0.05). (ii) Bioinformatics analysis revealed a putative conserved ‘canonical’ palmitoylation site in the juxtamembrane region of R1(124) and R2(135), plus additional non-conserved sites. This was confirmed using mutagenesis/acylbiotinyl exchange chemistry. Palmitoylation of R1(124) or R2(135) was shown to be required for efficient CSE of R1 and R2 and coupling to downstream signalling effectors including AMPK, AKT and ERK in cell lines and in vivo in mouse skeletal muscle respectively (all p < 0.05). These results provide increasing evidence that CSE of R1 and R2 is under complex, differential regulation and is essential for coupling adiponectin to downstream signalling effectors. Further studies are required to elaborate the detailed molecular mechanisms that may provide novel therapeutic opportunities.",
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Characterisation of the adiponectin receptors: Cell-surface expression and signal transduction of AdipoR1 and AdipoR2 are palmitoylation-dependent. / Keshvari, Sahar; Henstridge, Darren; Adams, Mark; O'Neill, Hayley M.; Hooper, John ; Febbraio, Mark A.; Whitehead, Jonathan P.

In: Obesity Research and Clinical Practice, Vol. 8, No. Suppl 1, 12.2014, p. 114-115.

Research output: Contribution to journalMeeting AbstractResearchpeer-review

TY - JOUR

T1 - Characterisation of the adiponectin receptors: Cell-surface expression and signal transduction of AdipoR1 and AdipoR2 are palmitoylation-dependent

AU - Keshvari, Sahar

AU - Henstridge, Darren

AU - Adams, Mark

AU - O'Neill, Hayley M.

AU - Hooper, John

AU - Febbraio, Mark A.

AU - Whitehead, Jonathan P

PY - 2014/12

Y1 - 2014/12

N2 - The adiponectin axis is a major regulator of metabolic, cardiovascular and inflammatory tone. Current understanding of the adiponectin receptors, R1 and R2, is rudimentary constraining our ability to target these atypical seven transmembrane receptors. Hence, we have started to characterise R1 and R2.We previously reported that R1 exhibits robust (60%) cell-surface expression (CSE) under steady-state conditions (no serum starvation) whilst R2 is undetectable and showed these differences were predicated by the non-conserved N-terminal, intracellular trunks [1]. Here we hypothesised that (i) CSE of R1/R2 is regulated, (ii) R1/R2 are subject to palmitoylation and this regulates CSE/function (as in GPCRs). To address these hypotheses we have employed bioinformatics, cell-based and in vivo approaches. We demonstrate that (i) serum starvation increases R2 CSE and adiponectin stimulates internalisation of R1/R2 (all p < 0.05). (ii) Bioinformatics analysis revealed a putative conserved ‘canonical’ palmitoylation site in the juxtamembrane region of R1(124) and R2(135), plus additional non-conserved sites. This was confirmed using mutagenesis/acylbiotinyl exchange chemistry. Palmitoylation of R1(124) or R2(135) was shown to be required for efficient CSE of R1 and R2 and coupling to downstream signalling effectors including AMPK, AKT and ERK in cell lines and in vivo in mouse skeletal muscle respectively (all p < 0.05). These results provide increasing evidence that CSE of R1 and R2 is under complex, differential regulation and is essential for coupling adiponectin to downstream signalling effectors. Further studies are required to elaborate the detailed molecular mechanisms that may provide novel therapeutic opportunities.

AB - The adiponectin axis is a major regulator of metabolic, cardiovascular and inflammatory tone. Current understanding of the adiponectin receptors, R1 and R2, is rudimentary constraining our ability to target these atypical seven transmembrane receptors. Hence, we have started to characterise R1 and R2.We previously reported that R1 exhibits robust (60%) cell-surface expression (CSE) under steady-state conditions (no serum starvation) whilst R2 is undetectable and showed these differences were predicated by the non-conserved N-terminal, intracellular trunks [1]. Here we hypothesised that (i) CSE of R1/R2 is regulated, (ii) R1/R2 are subject to palmitoylation and this regulates CSE/function (as in GPCRs). To address these hypotheses we have employed bioinformatics, cell-based and in vivo approaches. We demonstrate that (i) serum starvation increases R2 CSE and adiponectin stimulates internalisation of R1/R2 (all p < 0.05). (ii) Bioinformatics analysis revealed a putative conserved ‘canonical’ palmitoylation site in the juxtamembrane region of R1(124) and R2(135), plus additional non-conserved sites. This was confirmed using mutagenesis/acylbiotinyl exchange chemistry. Palmitoylation of R1(124) or R2(135) was shown to be required for efficient CSE of R1 and R2 and coupling to downstream signalling effectors including AMPK, AKT and ERK in cell lines and in vivo in mouse skeletal muscle respectively (all p < 0.05). These results provide increasing evidence that CSE of R1 and R2 is under complex, differential regulation and is essential for coupling adiponectin to downstream signalling effectors. Further studies are required to elaborate the detailed molecular mechanisms that may provide novel therapeutic opportunities.

U2 - 10.1016/j.orcp.2014.10.206

DO - 10.1016/j.orcp.2014.10.206

M3 - Meeting Abstract

VL - 8

SP - 114

EP - 115

JO - Obesity Research and Clinical Practice

JF - Obesity Research and Clinical Practice

SN - 1871-403X

IS - Suppl 1

ER -