Failed export of the adrenocorticotrophin receptor from the endoplasmic reticulum in non-adrenal cells: evidence in support of a requirement for a specific adrenal accessory factor
Open accessAbstract
Difficulty in expressing the adrenocorticotrophin (ACTH) receptor (melanocortin 2 receptor; MC2R) after transfection of various MC2R expression vectors has been experienced by many researchers. Reproducible evidence for expression has been obtained only in the Y6/OS3 corticoadrenal cell lines or in cells expressing endogenous melanocortin receptors. In order to determine the cause of this failure of expression we have undertaken the following studies. An MC2R expression plasmid was constructed in which the green fluorescent protein (GFP) coding region had been added to the C-terminus of the mature protein. Transfection of this plasmid into Y6 cells with a cAMP-responsive reporter plasmid demonstrated normal function of this receptor. Imaging of CHO cells expressing MC2R-GFP revealed perinuclear expression, although a cholecystokinin receptor (CCKR)-GFP construct was efficiently expressed at the cell surface. Y6 cells, in contrast, showed cell surface fluorescence after transfection with MC2R-GFP. Several other cell types showed a similar pattern of GFP distribution characteristic of retention in the endoplasmic reticulum. Counterstaining with an anti-KDEL antibody confirmed this location. Co-expression of the MC2R and the CCKR-GFP did not impair CCKR trafficking to the cell surface, implying a receptor-specific impairment to trafficking in the CHO cell which was absent in the Y6 cell.
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MC2R cannot function alone as an ACTH receptor.
“The MC2R cannot function alone as an ACTH receptor, which led to many difficulties in its characterization after initial cloning (33).”
From ACTH Antagonists (2016) · cites this paper as [33]
The Introduction and Discussion report that MC2R was difficult to characterize after cloning because functional expression was difficult to obtain in non-adrenal cells. The Discussion hypothesizes that MC2R requires an accessory factor for expression, but does not establish the categorical claim that it cannot function alone as an ACTH receptor. Page numbers were not available in our copy. (LLM, conf=0.86)
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“The MC2R cannot function alone as an ACTH receptor, which led to many difficulties in its characterization after initial cloning (33).”
BackgroundSelf-citationFridmanis, D., Petrovska, R., Kalnina, I., Slaidina, M., Peculis, R., Schiöth, H. B., & Klovins, J. (2010) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2010.02.032 · Cited by 30 · see citing works
- Authors
- Noon, L., Franklin, J., King, P., Goulding, N., Hunyady, L., & Clark, A.
- Affiliations
- University of London (GB)
- Semmelweis University (HU)
- Corresponding author
- Clark, AJ
- ORCID
- Hunyady, L
- Venue
- Journal of Endocrinology
- Publisher
- Bioscientifica
- Published
- 2002-07-01
- Volume
- 174
- Issue
- 1
- Pages
- 17-25
- ISSN
- 0022-0795 (ISSN-L)
- 1479-6805
- OpenAlex
- W2106099541
- Cited by
- 88
- References
- 44
- Citations by year
- 2026: 1 · 2025: 3 · 2023: 5 · 2022: 5 · 2021: 2 · 2020: 1 · 2019: 4 · 2017: 5 · 2016: 4 · 2015: 2 · 2014: 8 · 2013: 4 · 2012: 1
- FWCI
- 1.58
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Endocrinology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Queen Mary University of London
- University of Toronto
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Keywords
- melanocortin 2 receptor, endoplasmic reticulum retention, receptor trafficking, KDEL, cholecystokinin receptors, GFP fusion constructs, CHO cells
- MeSH terms
- Adrenocorticotropic Hormone, Animals, Endoplasmic Reticulum, Cricetinae, Luminescent Proteins, Receptors, Cholecystokinin, Transfection, Blotting, Western, CHO Cells, Receptors, Corticotropin, Receptor, Melanocortin, Type 2, Green Fluorescent Proteins
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