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

Sun Kyung Kim, Lindsey Barron, Cynthia S. Hinck, Elyse M. Petrunak, Kristin E. Cano, Avinash Thangirala, Brian Iskra, Molly Brothers, Machell Vonberg, Belinda Leal, Blair Richter, Ravindra Kodali, Alexander B. Taylor, Shoucheng Du, Christopher O. Barnes, Traian Sulea, Guillermo Calero, P. John Hart, Matthew J Hart, Borries Demeler & 1 others Andrew P. Hinck

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The transforming growth factorβ 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.

Original languageEnglish (US)
Pages (from-to)7173-7188
Number of pages16
JournalJournal of Biological Chemistry
Volume292
Issue number17
DOIs
StatePublished - Apr 28 2017

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Transforming Growth Factors
Monomers
Growth Factor Receptors
Crystallography
Aptitude
Heel
Immune system
Luciferases
Extracellular Matrix
Disease Progression
Immune System
Assays
Protein Isoforms
Fibrosis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kim, S. K., Barron, L., Hinck, C. S., Petrunak, E. M., Cano, K. E., Thangirala, A., ... Hinck, A. P. (2017). An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling. Journal of Biological Chemistry, 292(17), 7173-7188. https://doi.org/10.1074/jbc.M116.768754

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

In: Journal of Biological Chemistry, Vol. 292, No. 17, 28.04.2017, p. 7173-7188.

Research output: Contribution to journalArticle

Kim, SK, Barron, L, Hinck, CS, Petrunak, EM, Cano, KE, Thangirala, A, Iskra, B, Brothers, M, Vonberg, M, Leal, B, Richter, B, Kodali, R, Taylor, AB, Du, S, Barnes, CO, Sulea, T, Calero, G, Hart, PJ, Hart, MJ, Demeler, B & Hinck, AP 2017, 'An engineered transforming growth factor β (TGF-β) monomer that functions as a dominant negative to block TGF-β signaling', Journal of Biological Chemistry, vol. 292, no. 17, pp. 7173-7188. https://doi.org/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. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 17. pp. 7173-7188.
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AU - Cano, Kristin E.

AU - Thangirala, Avinash

AU - Iskra, Brian

AU - Brothers, Molly

AU - Vonberg, Machell

AU - Leal, Belinda

AU - Richter, Blair

AU - Kodali, Ravindra

AU - Taylor, Alexander B.

AU - Du, Shoucheng

AU - Barnes, Christopher O.

AU - Sulea, Traian

AU - Calero, Guillermo

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AU - Hart, Matthew J

AU - Demeler, Borries

AU - Hinck, Andrew P.

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