Mechanistic basis for the potent anti-angiogenic activity of Semaphorin 3f

Hou Fu Guo, Xiaobo Li, Matthew W. Parker, Johannes Waltenberger, Patrice M. Becker, Craig W. Vander Kooi

Research output: Contribution to journalArticle

Abstract

Neuropilin-1 (Nrp1), an essential type I transmembrane receptor, binds two secreted ligand families, vascular endothelial growth factor (VEGF) and class III Semaphorin (Sema3). VEGF-A and Sema3F have opposing roles in regulating Nrp1 vascular function in angiogenesis. VEGF-A functions as one of the most potent pro-angiogenic cytokines, while Sema3F is a uniquely potent endogenous angiogenesis inhibitor. Sema3 family members require proteolytic processing by furin to allow competitive binding to Nrp1. We demonstrate that the furin-processed C-terminal domain of Sema3F (C-furSema) potently inhibits VEGF-A-dependent activation of endothelial cells. We find that this potent activity is due to unique heterobivalent engagement of Nrp1 by two distinct sites in the C-terminal domain of Sema3F. One of the sites is the C-terminal arginine, liberated by furin cleavage, and the other is a novel upstream helical motif centered on the intermolecular disulfide. Using a novel chimeric C-furSema, we demonstrate that combining a single C-terminal arginine with the helical motif is necessary and sufficient for potent inhibition of binding of VEGF-A to Nrp1. We further demonstrate that the multiple furin-processed variants of Sema3A, with the altered proximity of the two binding motifs, have dramatically different potencies. This suggests that furin processing not only switches Sema3 to an activated form but also, depending on the site processed, can also tune potency. These data establish the basis for potent competitive binding of Sema3 to Nrp1 and provide a basis for the design of bivalent Nrp inhibitors.

Original languageEnglish (US)
Pages (from-to)7551-7558
Number of pages8
JournalBiochemistry®
Volume52
Issue number43
DOIs
StatePublished - Oct 29 2013

Fingerprint

Neuropilin-1
Semaphorins
Furin
Vascular Endothelial Growth Factor A
Competitive Binding
Arginine
Semaphorin-3A
Angiogenesis Inhibitors
Endothelial cells
Processing
Disulfides
Blood Vessels
Endothelial Cells
Chemical activation
Switches
Cytokines
Ligands

ASJC Scopus subject areas

  • Biochemistry

Cite this

Guo, H. F., Li, X., Parker, M. W., Waltenberger, J., Becker, P. M., & Vander Kooi, C. W. (2013). Mechanistic basis for the potent anti-angiogenic activity of Semaphorin 3f. Biochemistry®, 52(43), 7551-7558. https://doi.org/10.1021/bi401034q

Mechanistic basis for the potent anti-angiogenic activity of Semaphorin 3f. / Guo, Hou Fu; Li, Xiaobo; Parker, Matthew W.; Waltenberger, Johannes; Becker, Patrice M.; Vander Kooi, Craig W.

In: Biochemistry®, Vol. 52, No. 43, 29.10.2013, p. 7551-7558.

Research output: Contribution to journalArticle

Guo, HF, Li, X, Parker, MW, Waltenberger, J, Becker, PM & Vander Kooi, CW 2013, 'Mechanistic basis for the potent anti-angiogenic activity of Semaphorin 3f', Biochemistry®, vol. 52, no. 43, pp. 7551-7558. https://doi.org/10.1021/bi401034q
Guo HF, Li X, Parker MW, Waltenberger J, Becker PM, Vander Kooi CW. Mechanistic basis for the potent anti-angiogenic activity of Semaphorin 3f. Biochemistry®. 2013 Oct 29;52(43):7551-7558. https://doi.org/10.1021/bi401034q
Guo, Hou Fu ; Li, Xiaobo ; Parker, Matthew W. ; Waltenberger, Johannes ; Becker, Patrice M. ; Vander Kooi, Craig W. / Mechanistic basis for the potent anti-angiogenic activity of Semaphorin 3f. In: Biochemistry®. 2013 ; Vol. 52, No. 43. pp. 7551-7558.
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