N-glycosylation and expression in human tissues of the orphan GPR61 receptor

Paweł Kozielewicz, Hatun Alomar, Syaratul Dalina Yusoff, Gillian Grafton, Alison J. Cooper, S. John Curnow, James W. Ironside, Hardev Pall, Nicholas M. Barnes

Research output: Contribution to journalArticle

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Abstract

A number of members of the G protein-coupled receptor class of cell surface receptors are ‘orphans’ with no known endogenous ligand. One of these orphan receptors is GPR61; there are little data about its expression in human cells and tissues. In this study, we investigated the post-translational modification of GPR61 by N-glycosylation at an identified consensus N-glycosylation site (N12) and the impact of this modification upon the subcellular expression of the protein. The N-glycosylation inhibitor tunicamycin reduced the apparent molecular weight of immunoreactivity associated with myc-tagged GPR61 by 1–2 kDa, which was comparable to the evident molecular weight of the myc-tagged N12S GPR61 mutant with disrupted consensus N-glycosylation site. Analysis of GPR61 expression demonstrated that tunicamycin treatment reduced considerably heterologous expression of GPR61 in the cell membrane despite the N12S GPR61 mutant being readily expressed at the cell surface. These results demonstrate that GPR61 is subject to N-glycosylation but suggest this is not a prerequisite for cell surface expression, although N-glycosylation of other proteins may be important for cell membrane expression of GPR61. Expression of GPR61 protein was demonstrated at the cellular level in human hippocampus and human peripheral blood mononuclear cells. In the latter, there was a significantly higher expression of GPR61 in the Th17 cell subset in comparison with resting CD4+ cells, which may point toward a potential role for the GPR61 receptor in autoimmune diseases. This is the first report that GPR61 protein is subject to post-translational modification and is expressed in immune cell subsets and the hippocampus. These findings will help guide studies to investigate the function of GPR61.

Original languageEnglish
Pages (from-to)1982-1993
Number of pages12
JournalFEBS Open Bio
Volume7
Issue number12
DOIs
Publication statusPublished - 1 Dec 2017

Fingerprint

Glycosylation
Orphaned Children
Tissue
Tunicamycin
Cell membranes
Post Translational Protein Processing
Hippocampus
Proteins
Molecular Weight
Molecular weight
Cell Membrane
Th17 Cells
Cell Surface Receptors
G-Protein-Coupled Receptors
Autoimmune Diseases
Blood Cells
Blood
Cells
Ligands

Keywords

  • GPCR
  • hippocampus
  • lymphocyte
  • membrane trafficking
  • N-linked glycosylation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Kozielewicz, P., Alomar, H., Yusoff, S. D., Grafton, G., Cooper, A. J., Curnow, S. J., ... Barnes, N. M. (2017). N-glycosylation and expression in human tissues of the orphan GPR61 receptor. FEBS Open Bio, 7(12), 1982-1993. https://doi.org/10.1002/2211-5463.12339

N-glycosylation and expression in human tissues of the orphan GPR61 receptor. / Kozielewicz, Paweł; Alomar, Hatun; Yusoff, Syaratul Dalina; Grafton, Gillian; Cooper, Alison J.; Curnow, S. John; Ironside, James W.; Pall, Hardev; Barnes, Nicholas M.

In: FEBS Open Bio, Vol. 7, No. 12, 01.12.2017, p. 1982-1993.

Research output: Contribution to journalArticle

Kozielewicz, P, Alomar, H, Yusoff, SD, Grafton, G, Cooper, AJ, Curnow, SJ, Ironside, JW, Pall, H & Barnes, NM 2017, 'N-glycosylation and expression in human tissues of the orphan GPR61 receptor', FEBS Open Bio, vol. 7, no. 12, pp. 1982-1993. https://doi.org/10.1002/2211-5463.12339
Kozielewicz P, Alomar H, Yusoff SD, Grafton G, Cooper AJ, Curnow SJ et al. N-glycosylation and expression in human tissues of the orphan GPR61 receptor. FEBS Open Bio. 2017 Dec 1;7(12):1982-1993. https://doi.org/10.1002/2211-5463.12339
Kozielewicz, Paweł ; Alomar, Hatun ; Yusoff, Syaratul Dalina ; Grafton, Gillian ; Cooper, Alison J. ; Curnow, S. John ; Ironside, James W. ; Pall, Hardev ; Barnes, Nicholas M. / N-glycosylation and expression in human tissues of the orphan GPR61 receptor. In: FEBS Open Bio. 2017 ; Vol. 7, No. 12. pp. 1982-1993.
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