Effect of His-Gly-Lys Motif Derived from Domain 5 of High Molecular Weight Kininogen on Suppression of Cancer Metastasis Both in Vitro and in Vivo

Abstract

, an octapeptide including the HGK motif derived from D5 H , and HGK, a tripeptide, inhibited both cell adhesion and invasion in vitro. However, an octapeptide designated P-5m (K487R), in which Lys 487 was changed to Arg, did not inhibit either cell adhesion or invasion, and peptides HGR and HGG also had no inhibitory effect. Recombinant GST-D5 H expressed in Escherichia coli had a stronger inhibitory effect on cell adhesion and invasion in vitro than did GST-D5 H (K487R) in which Lys 487 was changed to Arg. Furthermore, P-5m (Gly 484 -Lys 491 ) peptide clearly suppressed lung metastasis in mice experimentally induced by using B16-F10 cells, but P-5m (G487R) had no effect. These data strongly indicate that both the HGK motif and lysine residue (Lys 487 ) play essential roles in inhibition of cell adhesion and invasion in vitro and in prevention of metastasis of cancer cells in vivo. We tried to identify the HGK motif binding protein on the surface of cancer cells. A 95-kDa surface biotin-labeled membrane protein was specifically detached from GST-D5 H by P-5 (His 479 -Lys 493 ) peptide but not by P-1 (Gly 402 -Lys 420 ) peptide originating from the N-terminal region of D5 H .

Identification of differentially expressed genes in clear cell renal cell carcinoma by analysis of full-length enriched cDNA library

“…For example, the up-expression of cyclin D3 gene in liver metastasis of colorectal cancer [10], matrix metallopeptidase 7 in oral squamous cell carcinoma [11], and regulator of G-protein signaling 5 in RCC [12] were observed and they were also identified as up-regulated genes in this study. Furthermore, the association of the apoptosis-related gene kininogen 1 [13] and the cytoskeleton-related gene gelsolin [14] with cancer were previously reported and the down-regulation of kininogen 1 and gelsolin in ccRCC tissues was also found in this study. These observations indicated that the identified genes by the full-length cDNA library method might be related with ccRCC progression.…”

Chimeric Glutathione S-Transferases Containing Inserts of Kininogen Peptides

“…These results suggest that D5-13 was indeed exposed on the surface of GST and presented in a conformational state with considerably greater anti-endothelial cell activity compared with the free peptide in solution. This conformational restraint might significantly enhance the interactions of D5-13 with an anti-angiogenic binding site expressed on proliferating endothelial cells (16).…”

“…In this study, we presented the design of engineered GST proteins carrying inserts of the HGK-rich motif derived from a sequence within D5 of HK. HK D5 contains endothelial cell-binding sites and inhibits angiogenesis through its ability to cause apoptosis of proliferating endothelial cells and to inhibit endothelial cell proliferation and migration (6,10,(12)(13)(14)(15)(16). Moreover, HGK peptides were found to block tumor metastasis (6,10,(12)(13)(14)(15)(16).…”

“…HK D5 contains endothelial cell-binding sites and inhibits angiogenesis through its ability to cause apoptosis of proliferating endothelial cells and to inhibit endothelial cell proliferation and migration (6,10,(12)(13)(14)(15)(16). Moreover, HGK peptides were found to block tumor metastasis (6,10,(12)(13)(14)(15)(16). Although the D5 peptide region within the context of intact high molecular weight kininogen or recombinant D5 induces endothelial cell apoptosis with an IC 50 in the low nanomolar range, the isolated D5 peptides express at least 2 logs lower activity, presumably due to enhanced flexibility and a failure to maintain the appropriate conformation for meaningful interactions with endothelial cells (6,10,(12)(13)(14)(15)(16).…”

“…HK D5 contains endothelial cell-binding sites and inhibits angiogenesis through its ability to cause apoptosis of proliferating endothelial cells (6,12,13) and to inhibit endothelial cell proliferation and migration (14). Moreover, a histidine-glycine-lysine (HGK) motif derived from this domain blocks tumor metastasis (6,10,(12)(13)(14)(15)(16). Although the exact mechanism of HGK peptide action has not been delineated (6,10,(12)(13)(14)(15)(16), this motif nevertheless represents an attractive target for the design of antitumor peptide therapeutics/drugs.…”