Pefitdes derived from high molecular weight kininogen (hk) domains 3 and 5 inhibit endothelial cell proliferation and induce endothelial cell apoptosis

Jingchuan Zhang*, Jose Juarez, David Elliott Shaw, Andrew P. Mazar, Keith R. McCrae

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

We have recently reported that two-chain high molecular weight kininogen (HKa) inhibits endothelial cell proliferation, induces endothelial cell apoptosis and inhibits angiogenesis. Recombinant high molecular weight kininogen domain 5 expresses similar activity, inhibiting endothelial cell proliferation by 50% (IC5(]) at a concentration of -60 nM. To further define the regions within kininogen domain 5 responsible for these effects, we prepared overlapping 16 amino acid peptides (each overlapping by 8 amino acids) encompassing kininogen domain 5 (aa 384-509), and measured their ability to inhibit endothelial cell proliferation and induce endothelial cell apoptosis. A similar strategy was used to assess potentially active regions within HK domain 3, which also contains regions that mediate binding of HK to endothelial cells. The most potent domain 5-derived peptides were found in the C-terminal region of the domain. H5-13 (KHGHGHGKHKNKGKKN; aa 480-495 of HK) inhibited endothelial cell proliferation by 86 ±13% at a concentration of 50 μM (IC!0 - 8 |iM), while H5-14 (HKNKGKKNGKHNGWKT; aa 488-503 of HK), used at the same concentration, inhibited proliferation by 93 ±7% (IC30 - 14 |iM). As observed with HKa and recombinant domain 5, the peptides caused endothelial cell apoptosis, and their activity was enhanced in the presence of Zn2. These results differ from a previous report, in which the domain 5 peptide most active in inhibiting endothelial cell proliferation was found to encompass amino acids 440-455 of HK. In addition, we also observed that two 16 amino acid peptides from HK domain 3 (H3-6, aa 267-282 of HK; H3-7, aa 275-290 of HK) also inhibited endothelial cell proliferation (ICJO - 1 nM) and induced apoptosis, though they were not freely soluble in aqueous buffers. Exogenous Zn2+ did not significantly affect the activity of the latter peptides. Neither the domain 3 or domain 5 peptides affected the proliferation of a number of other cell types, including primary cultures of fibroblasts and smooth muscle cells. These studies define active regions within HK domains 3 and 5 that inhibit endothelial cell proliferation and induce endothelial cell apoptosis. Current studies are focused on evaluating the antiangiogenic activity of these peptides in vitro.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART I
StatePublished - Dec 1 2000

ASJC Scopus subject areas

  • Hematology

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