Authors
Peng, Yi; Cao, Shufen; Kiselar, Janna; Xiao, Xiangzhu; Du, Zhanwen; Hsieh, An; Ko, Soobin; Chen, Yinghua; Agrawal, Prashansa; Zheng, Wenwei; Shi, Wuxian; Jiang, Wei; Yang, Lin; Chance, Mark R; Surewicz, Witold K; Buck, Matthias; Yang, Sichun
Abstract
The N-terminal transactivation domain (NTD) of estrogen receptor alpha, a well-known member of the family of intrinsically disordered proteins, mediates the receptor’s transactivation function. However, an accurate molecular dissection of NTD’s structure-function relationships remains elusive. Here, we show that the NTD adopts a mostly disordered, unexpectedly compact conformation that undergoes structural expansion on chemical denaturation. By combining small-angle X-ray scattering, hydroxyl radical protein footprinting, and computational modeling, we derive the ensemble-structures of the NTD and determine its ensemble-contact map revealing metastable long-range contacts, e.g., between residues I33 and S118. We show that mutation at S118, a known phosphorylation site, promotes conformational changes and increases coactivator binding. We further demonstrate via fluorine-19 (F-19) nuclear magnetic resonance that mutations near I33 alter F-19 chemical shifts at S118, confirming the proposed I33-S118 contact in the ensemble of structural disorder. These findings extend our understanding of how specific contact metastability mediates critical functions of disordered proteins.