Antiplasmin Activity of a Peptide That Binds to the Receptor-binding Site of Angiogenin*

It has been suggested that angiogenin binds to an actin-like molecule present on the surface of endothelial cells. Actin inhibits plasmin activity, but the angiogenin-actin complex is not active. In this report, we found that plasmin inhibits the interaction between angiogenin and actin suggesting a possibility that both angiogenin and plasmin may bind to a similar site on actin. Here we report that chANG, an antiangiogenin peptide that binds to the actin-binding site of angiogenin, inhibits the proteolytic activity of plasmin without any apparent effect on the activities of plasminogen activators and matrix metalloproteases. Its antiplasmin activity is comparable with that of actin. chANG inhibits plasmin activity via its binding to plasmin kringle domains while scrambled chANG does not bind to plasmin. chANG also inhibits the invasion of angiogenin-secreting human fibrosarcoma and colorectal carcinoma cells without effecting migration. Furthermore, chANG blocks angiogenesis induced by fibrosarcoma cells and metastasis of colorectal carcinoma cells to the liver. Therefore, the 11-amino acid peptide chANG has both antiangiogenin and antiplasmin activity, and could be useful in the development of anticancer agents.

[1]  G. Hu,et al.  An angiogenin-binding protein from endothelial cells. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[2]  T. Hallahan,et al.  Dual site model for the organogenic activity of angiogenin. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[3]  J. L. Bethune,et al.  Isolation and characterization of angiogenin, an angiogenic protein from human carcinoma cells. , 1985, Biochemistry.

[4]  C. Smith,et al.  Actin accelerates plasmin generation by tissue plasminogen activator. , 1991, The Journal of biological chemistry.

[5]  A. Gulino,et al.  Identification of plasminogen in Matrigel and its activation by reconstitution of this basement membrane extract. , 1996, BioTechniques.

[6]  Harper Jw,et al.  A covalent angiogenin/ribonuclease hybrid with a fourth disulfide bond generated by regional mutagenesis. , 1989 .

[7]  G. Hu,et al.  Angiogenin enhances actin acceleration of plasminogen activation. , 1993, Biochemical and biophysical research communications.

[8]  G. Hu,et al.  Actin is a binding protein for angiogenin. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. Riordan,et al.  Nuclear translocation of angiogenin in proliferating endothelial cells is essential to its angiogenic activity. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[10]  C Longstaff,et al.  Studies on the mechanisms of action of aprotinin and tranexamic acid as plasmin inhibitors and antifibrinolytic agents. , 1994, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[11]  B. Wiman,et al.  On the specific interaction between the lysine-binding sites in plasmin and complementary sites in alpha2-antiplasmin and in fibrinogen. , 1979, Biochimica et biophysica acta.

[12]  L. Zanetta,et al.  Control of type IV collagenase activity by components of the urokinase–plasmin system: a regulatory mechanism with cell‐bound reactants , 1997, The EMBO journal.

[13]  I. Brooks,et al.  Determination of rate and equilibrium binding constants for macromolecular interactions using surface plasmon resonance: use of nonlinear least squares analysis methods. , 1993, Analytical biochemistry.

[14]  H. Sato,et al.  Proteolytic activation of the precursor of membrane type 1 matrix metalloproteinase by human plasmin , 1997, FEBS letters.

[15]  Y. Gho,et al.  Anti-angiogenin Activity of the Peptides Complementary to the Receptor-binding Site of Angiogenin* , 1997, The Journal of Biological Chemistry.

[16]  K. Kohno,et al.  Modulation by bovine angiogenin of tubular morphogenesis and expression of plasminogen activator in bovine endothelial cells. , 1995, Biochemical and biophysical research communications.

[17]  S. Rosenberg,et al.  Isolation and characterization of actin and actin-binding protein from human platelets , 1981, The Journal of cell biology.

[18]  K. Danø,et al.  Plasminogen activators, tissue degradation, and cancer. , 1985, Advances in cancer research.

[19]  B. Vallee,et al.  Angiogenin activates endothelial cell phospholipase C. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[20]  J. Beintema,et al.  The amino acid sequence of human pancreatic ribonuclease. , 1984, Analytical biochemistry.

[21]  Brygida Berse,et al.  Vascular permeability factor (VPF, VEGF) in tumor biology , 1993, Cancer and Metastasis Reviews.

[22]  G. Hu,et al.  Angiogenin promotes invasiveness of cultured endothelial cells by stimulation of cell-associated proteolytic activities. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[23]  F. Soncin,et al.  Specific binding of angiogenin to calf pulmonary artery endothelial cells. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Magnus Malmqvist,et al.  Biospecific interaction analysis using biosensor technology , 1993, Nature.

[25]  B. Vallee,et al.  Actin is a surface component of calf pulmonary artery endothelial cells in culture. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Russ Granzow,et al.  Interactions in the Fourth Dimension , 1992, Bio/Technology.

[27]  Y Iwamoto,et al.  A rapid in vitro assay for quantitating the invasive potential of tumor cells. , 1987, Cancer research.

[28]  C. Smith,et al.  Actin is a noncompetitive plasmin inhibitor. , 1991, The Journal of biological chemistry.

[29]  R. Raines,et al.  Replacing a Surface Loop Endows Ribonuclease A with Angiogenic Activity (*) , 1995, The Journal of Biological Chemistry.

[30]  S. Takao,et al.  A modified invasion-3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay for quantitating tumor cell invasion. , 1994, Cancer research.

[31]  K. Kurachi,et al.  Sequence of the cDNA and gene for angiogenin, a human angiogenesis factor. , 1985, Biochemistry.

[32]  R. Dahiya,et al.  Mucin production by human colonic carcinoma cells correlates with their metastatic potential in animal models of colon cancer metastasis. , 1991, The Journal of clinical investigation.

[33]  B. Vallee,et al.  Angiogenin stimulates endothelial cell prostacyclin secretion by activation of phospholipase A2. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[34]  J. Riordan,et al.  Angiogenin activates phospholipase C and elicits a rapid incorporation of fatty acid into cholesterol esters in vascular smooth muscle cells. , 1990, Biochemistry.