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Updated: Mar 29, 2026

Fluorescence-Based Measurements of Phosphatidylserine/Phosphatidylinositol 4-Phosphate Exchange Between Membranes
Published on: March 14, 2021
Expression and purification of Steroidogenic Factor-1 (NR5A1) nuclear receptor ligand binding domain complexed with
Alexis N Campbell1, Alexander J Cutright2, Jessica Martin2
1Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, United States.
Abstract:
Steroidogenic Factor-1 (SF-1, NR5A1) is an orphan nuclear receptor that plays a critical role in proper development and adult function of the adrenals, gonads and the ventral medial hypothalamus in the brain. Structural studies have revealed that the SF-1 ligand-binding domain is capable of interaction with glycerophospholipids, such as phosphoinositides, in its binding pocket. Of these, the phosphoinositide PI(3,4,5)P3 (PIP3) has a high affinity for SF-1 and functions as a key regulatory ligand. This paper details the methods for the recombinant expression and purification of the SF-1 protein, as well as a novel protocol for isolating pure SF-1 bound to PIP3. These methods are a foundational step for future biophysical assays, crystallography studies, and the development of targeted therapeutics for SF-1-dependent adrenocortical carcinoma.
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