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Title: An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling

Abstract

The transforming growth factor β isoforms, TGF-β1, -β2, and -β3, are small secreted homodimeric signaling proteins with essential roles in regulating the adaptive immune system and maintaining the extracellular matrix. However, dysregulation of the TGF-β pathway is responsible for promoting the progression of several human diseases, including cancer and fibrosis. Despite the known importance of TGF-βs in promoting disease progression, no inhibitors have been approved for use in humans. Herein, we describe an engineered TGF-β monomer, lacking the heel helix, a structural motif essential for binding the TGF-β type I receptor (TβRI) but dispensable for binding the other receptor required for TGF-β signaling, the TGF-β type II receptor (TβRII), as an alternative therapeutic modality for blocking TGF-β signaling in humans. As shown through binding studies and crystallography, the engineered monomer retained the same overall structure of native TGF-β monomers and bound TβRII in an identical manner. Cell-based luciferase assays showed that the engineered monomer functioned as a dominant negative to inhibit TGF-β signaling with a Ki of 20–70 nM. Investigation of the mechanism showed that the high affinity of the engineered monomer for TβRII, coupled with its reduced ability to non-covalently dimerize and its inability to bind and recruit TβRI,more » enabled it to bind endogenous TβRII but prevented it from binding and recruiting TβRI to form a signaling complex. Such engineered monomers provide a new avenue to probe and manipulate TGF-β signaling and may inform similar modifications of other TGF-β family members.« less

Authors:
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Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
OTHER U.S. STATESUNIVERSITYNCINIHAHA
OSTI Identifier:
1355043
Resource Type:
Journal Article
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 292; Journal Issue: 17; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
ENGLISH
Subject:
60 APPLIED LIFE SCIENCES

Citation Formats

Kim, Sun Kyung, Barron, Lindsey, Hinck, Cynthia S., Petrunak, Elyse M., Cano, Kristin E., Thangirala, Avinash, Iskra, Brian, Brothers, Molly, Vonberg, Machell, Leal, Belinda, Richter, Blair, Kodali, Ravindra, Taylor, Alexander B., Du, Shoucheng, Barnes, Christopher O., Sulea, Traian, Calero, Guillermo, Hart, P. John, Hart, Matthew J., Demeler, Borries, and Hinck, Andrew P. An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling. United States: N. p., 2017. Web. doi:10.1074/jbc.M116.768754.
Kim, Sun Kyung, Barron, Lindsey, Hinck, Cynthia S., Petrunak, Elyse M., Cano, Kristin E., Thangirala, Avinash, Iskra, Brian, Brothers, Molly, Vonberg, Machell, Leal, Belinda, Richter, Blair, Kodali, Ravindra, Taylor, Alexander B., Du, Shoucheng, Barnes, Christopher O., Sulea, Traian, Calero, Guillermo, Hart, P. John, Hart, Matthew J., Demeler, Borries, & Hinck, Andrew P. An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling. United States. doi:10.1074/jbc.M116.768754.
Kim, Sun Kyung, Barron, Lindsey, Hinck, Cynthia S., Petrunak, Elyse M., Cano, Kristin E., Thangirala, Avinash, Iskra, Brian, Brothers, Molly, Vonberg, Machell, Leal, Belinda, Richter, Blair, Kodali, Ravindra, Taylor, Alexander B., Du, Shoucheng, Barnes, Christopher O., Sulea, Traian, Calero, Guillermo, Hart, P. John, Hart, Matthew J., Demeler, Borries, and Hinck, Andrew P. Wed . "An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling". United States. doi:10.1074/jbc.M116.768754.
@article{osti_1355043,
title = {An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling},
author = {Kim, Sun Kyung and Barron, Lindsey and Hinck, Cynthia S. and Petrunak, Elyse M. and Cano, Kristin E. and Thangirala, Avinash and Iskra, Brian and Brothers, Molly and Vonberg, Machell and Leal, Belinda and Richter, Blair and Kodali, Ravindra and Taylor, Alexander B. and Du, Shoucheng and Barnes, Christopher O. and Sulea, Traian and Calero, Guillermo and Hart, P. John and Hart, Matthew J. and Demeler, Borries and Hinck, Andrew P.},
abstractNote = {The transforming growth factor β isoforms, TGF-β1, -β2, and -β3, are small secreted homodimeric signaling proteins with essential roles in regulating the adaptive immune system and maintaining the extracellular matrix. However, dysregulation of the TGF-β pathway is responsible for promoting the progression of several human diseases, including cancer and fibrosis. Despite the known importance of TGF-βs in promoting disease progression, no inhibitors have been approved for use in humans. Herein, we describe an engineered TGF-β monomer, lacking the heel helix, a structural motif essential for binding the TGF-β type I receptor (TβRI) but dispensable for binding the other receptor required for TGF-β signaling, the TGF-β type II receptor (TβRII), as an alternative therapeutic modality for blocking TGF-β signaling in humans. As shown through binding studies and crystallography, the engineered monomer retained the same overall structure of native TGF-β monomers and bound TβRII in an identical manner. Cell-based luciferase assays showed that the engineered monomer functioned as a dominant negative to inhibit TGF-β signaling with a Ki of 20–70 nM. Investigation of the mechanism showed that the high affinity of the engineered monomer for TβRII, coupled with its reduced ability to non-covalently dimerize and its inability to bind and recruit TβRI, enabled it to bind endogenous TβRII but prevented it from binding and recruiting TβRI to form a signaling complex. Such engineered monomers provide a new avenue to probe and manipulate TGF-β signaling and may inform similar modifications of other TGF-β family members.},
doi = {10.1074/jbc.M116.768754},
journal = {Journal of Biological Chemistry},
issn = {0021-9258},
number = 17,
volume = 292,
place = {United States},
year = {2017},
month = {2}
}

Works referenced in this record:

Overview of the CCP 4 suite and current developments
journal, March 2011

  • Winn, Martyn D.; Ballard, Charles C.; Cowtan, Kevin D.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
  • DOI: 10.1107/S0907444910045749

TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs: TGF-β signals through autonomous TβRI:TβRII pairs
journal, March 2011

  • Huang, Tao; David, Laurent; Mendoza, Valentín
  • The EMBO Journal, Vol. 30, Issue 7
  • DOI: 10.1038/emboj.2011.54

Ternary Complex of Transforming Growth Factor-β1 Reveals Isoform-specific Ligand Recognition and Receptor Recruitment in the Superfamily
journal, March 2010

  • Radaev, Sergei; Zou, Zhongcheng; Huang, Tao
  • Journal of Biological Chemistry, Vol. 285, Issue 19
  • DOI: 10.1074/jbc.M109.079921

Assembly of TβRI:TβRII:TGFβ Ternary Complex in vitro with Receptor Extracellular Domains is Cooperative and Isoform-dependent
journal, December 2005

  • Zúñiga, Jorge E.; Groppe, Jay C.; Cui, Yumin
  • Journal of Molecular Biology, Vol. 354, Issue 5
  • DOI: 10.1016/j.jmb.2005.10.014

XDS
journal, January 2010

  • Kabsch, Wolfgang
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
  • DOI: 10.1107/S0907444909047337

Mechanism of activation of the TGF-β receptor
journal, August 1994

  • Wrana, Jeffrey L.; Attisano, Liliana; Wieser, Rotraud
  • Nature, Vol. 370, Issue 6488
  • DOI: 10.1038/370341a0

Unchaining the beast; insights from structural and evolutionary studies on TGFβ secretion, sequestration, and activation
journal, August 2013


Cooperative Assembly of TGF-β Superfamily Signaling Complexes Is Mediated by Two Disparate Mechanisms and Distinct Modes of Receptor Binding
journal, February 2008


iMOSFLM : a new graphical interface for diffraction-image processing with MOSFLM
journal, March 2011

  • Battye, T. Geoff G.; Kontogiannis, Luke; Johnson, Owen
  • Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
  • DOI: 10.1107/S0907444910048675

NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy
journal, December 2014


Features and development of Coot
journal, March 2010

  • Emsley, P.; Lohkamp, B.; Scott, W. G.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
  • DOI: 10.1107/S0907444910007493

PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010

  • Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
  • DOI: 10.1107/S0907444909052925

Monte Carlo analysis of sedimentation experiments
journal, June 2007


Tgf-β Signal Transduction
journal, June 1998


NMRPipe: A multidimensional spectral processing system based on UNIX pipes
journal, November 1995

  • Delaglio, Frank; Grzesiek, Stephan; Vuister, GeertenW.
  • Journal of Biomolecular NMR, Vol. 6, Issue 3
  • DOI: 10.1007/BF00197809

Methods for the Design and Analysis of Sedimentation Velocity and Sedimentation Equilibrium Experiments with Proteins
journal, April 2010


Structural Biology and Evolution of the TGF-β Family
journal, September 2016

  • Hinck, Andrew P.; Mueller, Thomas D.; Springer, Timothy A.
  • Cold Spring Harbor Perspectives in Biology, Vol. 8, Issue 12
  • DOI: 10.1101/cshperspect.a022103

TGFβ in Cancer
journal, July 2008


Overexpression of human transforming growth factor-β1 using a recombinant CHO cell expression system
journal, October 2004


Phaser crystallographic software
journal, July 2007

  • McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
  • Journal of Applied Crystallography, Vol. 40, Issue 4
  • DOI: 10.1107/S0021889807021206