Identification of novel short peptides derived from the α4, α5, and α6 fibrils of type IV collagen with anti-angiogenic properties

doi:10.1016/j.bbrc.2006.12.231

Biochemical and Biophysical Research Communications

Volume 354, Issue 2, 9 March 2007, Pages 434-439

https://doi.org/10.1016/j.bbrc.2006.12.231 Get rights and content

Abstract

Angiogenesis, or neovascularization, is tightly controlled by positive and negative regulators, many of which reside in the extracellular matrix. We have now identified eight novel 19- to 20-residue peptides derived from the α4, α5, and α6 fibrils of type IV collagen, which we have designated tetrastatins, pentastatins, and hexastatins, respectively. We have shown that these endogenous peptides suppress the proliferation and migration of HUVECs in vitro. By performing clustering analyses of the sequences using sequence similarity criteria and of the experimental results using a hierarchical algorithm, we report that the clusters identified by the experimental results coincide with the sequence-based clusters, indicating a tight relationship between peptide sequence and anti-angiogenic potency. These peptides may have potential as anti-angiogenic therapeutic agents.

Section snippets

Materials and methods

Cell culture. Primary human umbilical vein endothelial cells (HUVECs) from a single donor were purchased from Cambrex (Walkersville, MD). The cells were propagated in EGM-2 medium, consisting of a basal cell medium with 2% FBS, growth factors (hbFGF and VEGF) and antibiotics (gentamicin/amphotericin B). All the cells used were from passage 3 to passage 6.

Peptide synthesis and handling. The peptides used were produced by the custom peptide synthesis facility in the Department of Oncology, Johns

Results and discussion

By using a bioinformatics approach, we have identified a set of eight novel short peptides, 19–20 amino acids in length, that are derived from the non-collagenous domains of the α4, α5, and α6 fibrils of type IV collagen: three derived from α4, three from α5, and two from α6 fibrils. Technical details of this bioinformatics analysis are not essential for the present study and are the subject of a separate report. These peptides display sequence similarities to known anti-angiogenic fragments

Conclusions

Using a bioinformatics approach, we have identified a set of short peptides derived from the α4, α5, and α6 fibrils of type IV collagen that have putative anti-angiogenic properties. We named these peptides tetrastatins, pentastatins, and hexastatins, respectively. Using HUVECs, we have evaluated the potency of these peptides as anti-angiogenic agents in proliferation and migration assays. Endothelial cell proliferation and migration are vital for the initiation and growth of a vascular sprout

Acknowledgments

The authors thank Zaver Bhujwalla and Roberto Pili for useful discussions; David Noren, Venu Raman, Kristine Glunde, Noriko Mori, Paul Winnard, and David Qian for their valuable advice on the experimental assays and Deborah McClellan for editorial assistance. The work was supported in part by NIH Grants NHLBI R01 HL079653 and NCI P50 CA103175.

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Identification of novel short peptides derived from the α4, α5, and α6 fibrils of type IV collagen with anti-angiogenic properties

Abstract

Section snippets

Materials and methods

Results and discussion

Conclusions

Acknowledgments

Int. J. Biochem. Cell. Biol.

Curr. Opin. Cell. Biol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Pharmacol. Toxicol. Methods

Tumor angiogenesis: therapeutic implications

N. Engl. J. Med.

Angiogenesis in life, disease and medicine

Nature