Expression, purification and crystallization of the ancestral androgen receptor-DHT complex

• Type of Material: Article
• Authors: Jennifer K. Colucci, Emory University; Eric Ortlund, Emory University
• Language: English
• Date: 2013-09-01
• Publisher: INT UNION CRYSTALLOGRAPHY
• Publication Version: Final Published Version
• Copyright Statement: © 2013 International Union of Crystallography All rights reserved.
• Final Published Version (URL): http://dx.doi.org/10.1107/S1744309113018745
• Title of Journal or Parent Work: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS
• Volume: 69
• Issue: 9
• Start Page: 994
• End Page: 996
• Grant/Funding Information: This research is supported by R01 GM081592 (Joseph Thornton PI, EAO coPI), start-up funds from Emory University (EAO) and JKC’s American Heart Association pre-doctoral fellowship (10PRE3530007).; Supporting institutions may be found at http://www.ser-cat.org/members.html.; Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. W-31-109-Eng-38.
• Keywords: Life Sciences & Biomedicine; PROTEIN; Biochemical Research Methods; Biophysics; PROSTATE-CANCER; Biochemistry & Molecular Biology; EVOLUTION; Physical Sciences; LESSONS; Crystallography; MODELS; PROGRESSION; Science & Technology
• Research Categories: Biology, Molecular; Chemistry, Biochemistry

Abstract

Steroid receptors (SRs) are a closely related family of ligand-dependent nuclear receptors that mediate the transcription of genes critical for development, reproduction and immunity. SR dysregulation has been implicated in cancer, inflammatory diseases and metabolic disorders. SRs bind their cognate hormone ligand with exquisite specificity, offering a unique system to study the evolution of molecular recognition. The SR family evolved from an estrogen-sensitive ancestor and diverged to become sensitive to progestagens, corticoids and, most recently, androgens. To understand the structural mechanisms driving the evolution of androgen responsiveness, the ancestral androgen receptor (ancAR1) was crystallized in complex with 5α- dihydrotestosterone (DHT) and a fragment of the transcriptional mediator/intermediary factor 2 (Tif2). Crystals diffracted to 2.1Å resolution and the resulting structure will permit a direct comparison with its progestagen-sensitive ancestor, ancestral steroid receptor 2 (AncSR2).