Purification and characterization of recombinant human mineralocorticoid receptor.

Molecular and Cellular Endocrinology
Colin D ClyneMorag J Young

Abstract

The mineralocorticoid receptor (MR) plays a critical role in the maintenance of electrolyte homeostasis and blood pressure via direct effects on the distal nephron and the cardiovascular system. The MR also has an important role in the pathology of cardiovascular disease, particularly heart failure, and is therefore an attractive therapeutic target. However, renal side effects limit its use in the clinic. Previous studies of MR molecular pharmacology have been performed on its isolated ligand-binding domain (LBD); however, current evidence suggests that nuclear receptor LBDs behave differently in isolation, than in the context of the full-length receptor. To date, technical issues have precluded production of full-length MR, thereby preventing molecular and structural studies of the MR LBD in its natural context. Here, we describe expression and purification of full-length human MR (hMR). hMR was expressed in Sf9 insect cells with an N-terminal biotinylated (bt)-tag, and stabilised by addition of ligand. bt-hMR exhibited ligand-binding and transactivation properties similar to that of the native protein. Affinity purification using an avidin matrix yielded approximately 120mug MR protein from 0.5lt Sf9 culture, and the receptor...Continue Reading

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Citations

May 16, 2009·Current Hypertension Reports·Frances McManusE Marie Freel
Jul 26, 2011·Molecular and Cellular Endocrinology·Jun Yang, Peter J Fuller
Feb 11, 2015·Molecular and Cellular Endocrinology·Peter J Fuller
Jan 4, 2015·Journal of Molecular Endocrinology·Jun YangMorag J Young
Jul 23, 2014·Endocrinology·Katelee Barrett MuellerIris Z Jaffe
Feb 26, 2010·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Monika Proszkowiec-Weglarz, Tom E Porter
Jul 17, 2015·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Jessica A ChadwickJill A Rafael-Fortney
Jul 8, 2014·Molecular Endocrinology·Jun YangMorag J Young
Aug 26, 2015·Molecular Endocrinology·Alicia M BlessingDaniel E Frigo
Sep 21, 2021·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Namita MahadikRajkumar Banerjee
Apr 27, 2010·Current Opinion in Endocrinology, Diabetes, and Obesity

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