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.

نویسندگان

  • L A Noon
  • J M Franklin
  • P J King
  • N J Goulding
  • L Hunyady
  • A J L Clark
چکیده

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.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

An evidence for a potassium channel in endoplasmic reticulum based on single channel recording in bilayer lipid membrane

Introduction Numerous studies have demonstrated the presence of potassium selective channels in membranes internal organelles. These channels are essential to a large variety of cellular processes including intracellular 2+ a signaling, protein recycling, charge neutralization and cell protection. In contrast to the sarcoplasmic reticulum + here potassium channels have been clearly ...

متن کامل

Allantoin improves methionine-choline deficient diet-induced nonalcoholic steatohepatitis in mice through involvement in endoplasmic reticulum stress and hepatocytes apoptosis-related genes expressions

Objective(s): Non-alcoholic steatohepatitis (NASH) is defined by steatosis and inflammation in the hepatocytes, which can progress to cirrhosis and possibly hepatocellular carcinoma. However, current treatments are not entirely effective. Allantoin is one of the principal compounds in many plants and an imidazoline I receptor agonist as well. Allantoin has positive eff...

متن کامل

Actors of necroptosis scenario in cell\'s scene

Necroptosis, as a novel concept, has been recently introduced in scientific literature. Much of our knowledge about necroptosis comes from ligation of tumor necrosis factor-α to its receptor, TNF receptor 1. Receptor-interacting protein kinase 1, receptor-interacting protein kinase 3 and its substrate, the pseudokinase mixed lineage kinase domain-like protein, have been comprehensively st...

متن کامل

Adrenal simple cyst in a young female patient: A case report

Introduction: Adrenal gland cysts with variable symptoms are rare cases of over-kidney clinical conditions. Adrenal cysts are accidentally identified, which is referred to as incidentaloma. Adrenal cysts are non-specific and show radiological findings and Detection of these cysts is usually random. In this article,  a patient with a simple adrenal cyst is introduced Case presentation: The p...

متن کامل

Cytoplasmic acidification reduces potassium channel activities in the endoplasmic reticulum of rat hepatocytes

Introduction: Intracellular pH (pHi) regulates essentially all aspects of cellular activities. However, it is unknown how endoplasmic reticulum (ER) potassium channels sense pHi. In this study, we investigate the direct effects of pHi on ER potassium channels. Methods: We used channel incorporation into the bilayer lipid membrane method. L-α-phosphatidylcholine, a membrane lipid, was extrac...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • The Journal of endocrinology

دوره 174 1  شماره 

صفحات  -

تاریخ انتشار 2002