Engineering synthetic antibody binders for allosteric inhibition of prolactin receptor signaling

Cell Communication and Signaling

Short Report

Somnath Mukherjee¹  Anthony A Kossiakoff¹  Erica M Duguid¹  Shahir S Rizk²  Anna Szymborska³  Jean-Louis K Kouadio⁴  Jiamao Zheng⁵  Charles V Clevenger⁶ 

[1] Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA;Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA;Current Address: Boler –Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, Notre Dame, IN, USA;Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA;Current Address: Max Delbrueck Center for Molecular Medicine, Berlin, Germany;Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA;Current Address: Monsanto Co. 700 Chesterfield Parkway, Chesterfield, MO, USA;Department of Pathology, Northwestern University, Chicago, IL, USA;Department of Pathology, Northwestern University, Chicago, IL, USA;Current Address: Department of Pathology, Virginia Commonwealth University, Richmond, VA, USA;

关键词: Prolactin signaling;    Synthetic antibody;    Phage display;    Allostery;   

DOI  :  10.1186/s12964-014-0080-8

 received in 2014-10-09, accepted in 2014-12-17,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundMany receptors function by binding to multiple ligands, each eliciting a distinct biological output. The extracellular domain of the human prolactin receptor (hPRL-R) uses an identical epitope to bind to both prolactin (hPRL) and growth hormone (hGH), yet little is known about how each hormone binding event triggers the appropriate response.FindingsHere, we utilized a phage display library to generate synthetic antibodies (sABs) that preferentially modulate hPRL-R function in a hormone-dependent fashion. We determined the crystal structure of a sAB-hPRL-R complex, which revealed a novel allosteric mechanism of antagonism, whereby the sAB traps the receptor in a conformation more suitable for hGH binding than hPRL. This was validated by examining the effect of the sABs on hormone internalization via the hPRL-R and its downstream signaling pathway.ConclusionsThe findings suggest that subtle structural changes in the extracellular domain of hPRL-R induced by each hormone determine the biological output triggered by hormone binding. We conclude that sABs generated by phage display selection can detect these subtle structural differences, and therefore can be used to dissect the structural basis of receptor-ligand specificity.

【 授权许可】

Unknown   
© Rizk et al.; licensee BioMed Central. 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.