Abstract
The angiotensin (Ang) IV analog norleual [Nle-Tyr-Leu-ψ-(CH2-NH2)3-4-His-Pro-Phe] exhibits structural homology with the hinge (linker) region of hepatocyte growth factor (HGF) and is hypothesized to act as a hinge region mimic. Norleual competitively inhibited the binding of HGF to its receptor c-Met in mouse liver membranes, with an IC50 value of 3 pM. Predictably, norleual was able to inhibit HGF-dependent signaling, proliferation, migration, and invasion in multiple cell types at concentrations in the picomolar range. Ex vivo studies demonstrated that norleual exhibited potent antiangiogenic activity, an attribute that would be predicted for a HGF/c-Met antagonist. Furthermore, norleual suppressed pulmonary colonization by B16-F10 murine melanoma cells, which are characterized by an overactive HGF/c-Met system. Together, these data suggest that AngIV analogs exert at least some of their biological activity through interference with the HGF/c-Met system and may have utility as therapeutic agents in disorders that are dependent on an intact HGF/c-Met system. Finally, the ability of norleual to induce marked biological responses in human embryonic kidney cells, which do not express insulin-responsive aminopeptidase (IRAP), coupled with the observed effects of norleual on the HGF/c-Met system, casts doubt on the physiological significance of AngIV-dependent inhibition of IRAP.
Footnotes
- Received September 22, 2009.
- Accepted January 15, 2010.
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B.J.Y. and P.D.E. contributed equally to this work.
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This work was supported by the state of Washington and Pacific Northwest Biotechnology, LLC and the National Institutes of Health National Eye Institute [Grant EY012836] (to M.D.V.).
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Portions of the work described here can be found in Yamamoto BJ (2006) Norleual, an Angiotensin IV Receptor Ligand and c-Met Antagonist, Ph.D. thesis, Washington State University, Pullman, WA and Elias JD (2008) On the Development of the Angiotensin IV Ligands, Norleual, and Nle′-angiotensin IV as Anticancer and Wound Healing Agents, Ph.D. thesis, Washington State University, Pullman, WA.
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.109.161711.
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↵ The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
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ABBREVIATIONS:
- AngIV
- angiotensin IV
- IRAP
- insulin-responsive aminopeptidase
- LTP
- long-term potentiation
- HGF
- hepatocyte growth factor
- norleual
- Nle-Tyr-Leu-ψ-(CH2-NH2)3-4-His-Pro-Phe
- Erk
- extracellular signal-regulated kinase
- EGM
- endothelial growth medium
- HEK
- human embryonic kidney
- MDCK
- Madin-Darby canine kidney
- DMEM
- Dulbecco's modified Eagle's medium
- FBS
- fetal bovine serum
- HUVEC
- human umbilical vein endothelial cell
- BSA
- bovine serum albumin
- PBS
- phosphate-buffered saline
- TBS
- Tris-buffered saline
- MTT
- 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan
- uPA
- urokinase-type plasminogen activator
- MSP
- macrophage-stimulating protein
- MEK
- mitogen-activated protein kinase kinase
- Gab1
- growth factor receptor bound protein-2-associated binder
- B428
- 4-iodo[b]thiophene-2-carboxamidine
- IP
- immunoprecipitated
- IB
- immunoblotted.
- Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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