Estrogen Receptor Folding Modulates cSrc Kinase SH2 Interaction via a Helical Binding Mode

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• 作者：Lidia Nieto ; Inga M. Tharun ; Mark Balk ; Hans Wienk ; Rolf Boelens ; Christian Ottmann ; Lech-Gustav Milroy ; Luc Brunsveld
• 刊名：ACS Chemical Biology
• 出版年：2015
• 出版时间：November 20, 2015
• 年：2015
• 卷：10
• 期：11
• 页码：2624-2632
• 全文大小：506K
• ISSN：1554-8937

文摘

The estrogen receptors (ERs) feature, next to their transcriptional role, important nongenomic signaling actions, with emerging clinical relevance. The Src Homology 2 (SH2) domain mediated interaction between cSrc kinase and ER plays a key role in this; however the molecular determinants of this interaction have not been elucidated. Here, we used phosphorylated ER peptide and semisynthetic protein constructs in a combined biochemical and structural study to, for the first time, provide a quantitative and structural characterization of the cSrc SH2鈥揈R interaction. Fluorescence polarization experiments delineated the SH2 binding motif in the ER sequence. Chemical shift perturbation analysis by nuclear magnetic resonance (NMR) together with molecular dynamics (MD) simulations allowed us to put forward a 3D model of the ER-SH2 interaction. The structural basis of this protein鈥損rotein interaction has been compared with that of the high affinity SH2 binding sequence GpYEEI. The ER features a different binding mode from that of the 鈥渢wo-pronged plug two-hole socket鈥?model in the so-called specificity determining region. This alternative binding mode is modulated via the folding of ER helix 12, a structural element directly C-terminal of the key phosphorylated tyrosine. The present findings provide novel molecular entries for understanding nongenomic ER signaling and targeting the corresponding disease states.