Angiostatin/Kringles and Related Products
 

Endostatin/Plasminogen and Related Products
 

M= Mouse; R=Rat; H=Human; Rb=Rabbit; G=goat; B=Bovine, MO=Monkey; P=pig; CT= near C-terminus; NT=near N-terminus; Internal=Middle of protein. EC=extracellular; CP=cytoplasmic domains *


Angiogenin, Angiostatin, Endostatin and Plasminogen Antibodies-General Information

Embryonic vascular system undergoes a series of complex, highly regulated series of events involving differentiation, migration and association of primitive endothelial cells. This process is termed vasculogenesis. A further remodeling of the primitive vascular system forms the mature cardiovascular system. This process is known as angiogenesis (sprouting of new capillary vessels from pre-existing vasculature). Angiogenesis accounts for the formation of vasculature into previously avascular organs such as brain and kidney. Angiogenic activity in the adult is required during the normal tissue repair, and for the remodeling of the female reproductive organs (ovulation and placental development). Certain pathological conditions, such as tumor growth and diabetic retinopathy, also require angiogenesis.

Recent studies have identified several proteolytic fragments or cryptic domains of proteins that act as inhibitors of angiogenesis. These include fragments of plasminogen such as Angiostatin protein (kringles 1-4) and kringles 1-5, C-terminal proteolytic fragment of Collagen XVIII (Endostatin protein), the NC10 domain of collagen 15 (Restin), the C-terminal hemopexin-like domain of MMP-2 (PEX), the N-terminal fragment of prolactin, and the N-terminally truncated platelet factor.

The 20 kDa fragment of Collagen XVIII known as Endostatin protein inhibits tumor progression and induction of endothelial cell apoptosis. Administration of Endostatin protein to mice bearing certain tumors caused tumor regression without the development of drug resistance. Endostatin protein also inhibited systemic angiogenesis, primary tumor growth, and the development of primary metastatic lesions. It also reduced the levels of antiapoptotic proteins Bcl-2 and Bcl-xL and an increase in apoptosis of endothelial cells in vitro.

Plasminogen prescursor or profibrinolysin or plasma trypsinogen (human 810 aa, chromosome 6q26) is synthesized in the kidney. It is present in plasma and many other extracellular fluids. Plasminogen is the zymogen in the circulating blood from which plasmin is formed. Plasminogen is a single-chain glycoprotein with 790 amino acid residues. Activation to the active form, plasmin, by urokinase involves cleavage at the arg-val bond between residues 560 and 561, resulting in the formation of the 2-chain plasmin molecule held together by 2 disulfide linkages. The heavier chain contains about 411 residues and the lighter chain about 233. The main function of plasmin is the digestion of fibrin in blood clots. Plasmin is a proteolytic enzyme with a specificity similar to that of trypsin. Like trypsin, plasmin belongs to the family of serine proteinases, in which the active site catalytic triad, his-57, asp-102, and ser-195 (chymotrypsin numbering), is situated in the light chain. The mature chain represents 20-810 aa. Processed active peptide domain is 20-97 aa. Angiostatin mature chain represents 98-810 aa. This regions contains 5 structurally related kringles domains K1-5 (Kringle 1, 103-181 aa; Kringle 2, 184-262 aa; Kringle 3, 275-352 aa; Kringle 4, 377-454 aa; Kringle 5, 481-560 aa). Each Kringle domain contains 6 conserved cysteines. Kringles 1-5 domains share approx 45-50% sequence homology.

Angiostatin protein, a proteolytic fragment of plasminogen, is comprised of the first four kringle regions. It prevents the growth of endothelial cells, and its systemic administration inhibits the growth of primary carcinomas in mice. Kringles 1-3 fragment has a greater inhibitory activity than the kringles 1-4 fragment. The protease-activated kringles 1-5 is the most potent plasminogen fragment with over 50-fold greater endothelial cell specific inhibitory activity. Its systemic administration inhibited the growth of fibrosarcoma and significantly reduced neovascularization.

Angiogenin is a single polypeptide chain of 123 aa (mol wt ~14 kDa) secreted by tumor cells. It is a potent inhibitor of neovascularization. It specifically binds to endothelial cells and elicits second messenger systems. It has ribonucleolytic activity and is 33% identical to pancreatic RNAse A. Angiogenin has also been shown to undergo nuclear translocation in endothelial cells via receptor-mediated endocytosis and nuclear localization sequence-assisted nuclear import. Human and mouse angiogenin are 76% identical. The critical catalytic residues of human angiogenin are conserved in the mouse protein, as are the six cysteines necessary for disulfide bond formation. Several C-terminal synthetic peptides, including (Ang 108-123), significantly decreases angiogenin-induced neovascularization. Angiogenin-related protein (Angrp), the putative product of a recently discovered mouse gene, shares 78% sequence identity with mouse angiogenin. Angrp, despite having greater ribonucleolytic activity than angiogenin, is not angiogenic.

rev. 50107A

List of publications using ADI's Antibodies to various Angiogenesis Related items

Product , Authors, Year of Pub, Journal, Western IHC, Comments, IP

Ang1
Yuan Hai Tao 2002 Angiopoietin correlates with glomerular capillary loss in anti-glomerular basement membrane glomerulonephritis Kidney International. 61(6):2078-2089 WB IHC mouse kidney

ang-1
Long, David A. 2001 Increased Renal Angiopoietin-1 Expression in Folic Acid-Induced Nephrotoxicity in Mice J Am Soc Nephrol 2001 12: 2721-2731 WB IHC mouse ang-1/2

ang-1
Zheng W 2004 DITPA stimulates arteriolar growth and modifies myocardial postinfarction remodeling Am J Physiol Heart Circ Physiol, 286: 1994 - 2000 WB rat

ang-1
(ang11) Krikun G 2002 Abnormal Uterine Bleeding during Progestin-Only Contraception May Result from Free Radical-Induced Alterations in Angiopoietin Expression Am. j. Pathol. 161, 979-986 WB IHC 4% PF/praffin embedded human endometrium/HESC cells

Ang-1/ang11
Hayashi, T 2003 Temporal Profile of Angiogenesis and Expression of Related Genes in the Brain After Ischemia. Journal of Cerebral Blood Flow & Metabolism. 23(2):166-180 IHC mouse tissues

ang-1m
Long, David A. 2001 Increased Renal Angiopoietin-1 Expression in Folic Acid-Induced Nephrotoxicity in Mice J Am Soc Nephrol 2001 12: 2721-2731 WB IHC mouse ang-1/2

ang2
Yuan Hai Tao 2002 Angiopoietin correlates with glomerular capillary loss in anti-glomerular basement membrane glomerulonephritis Kidney International. 61(6):2078-2089 WB IHC mouse kidney

ang-2
Zheng W 2004 DITPA stimulates arteriolar growth and modifies myocardial postinfarction remodeling Am J Physiol Heart Circ Physiol, 286: 1994 - 2000 WB rat

Ang-2/ang21
Hayashi, T 2003 Temporal Profile of Angiogenesis and Expression of Related Genes in the Brain After Ischemia. Journal of Cerebral Blood Flow & Metabolism. 23(2):166-180 IHC mouse tissues

ANST
Basile DP 2004 Angiostatin and matrix metalloprotease expression following ischemic acute renal failure Am J Physiol Renal Physiol, May 2004; 286: 893 - 902 WB, used pure anst as std

ANST
Hatziapostolou M 2003 Different inhibitors of plasmin differentially affect angiostatin production and angiogenesis European Journal of Pharmacology 460, 1-8 WB, chicken Chorioallantoic membrane tissues

FLK1/KDR
Sugano M 2004 Intramuscular Gene Transfer of Soluble Tumor Necrosis Factor- Receptor 1 Activates Vascular Endothelial Growth Factor Receptor and Accelerates Angiogenesis in a Rat Model of Hindlimb Ischemia Circulation. 109(6):797-802 WB IP
VEGF Recombinant human
Storment, John M. 2001 Estrogen augments the vasodilatory effects of vascular endothelial growth factor in the uterine circulation of the rat.[ American Journal of Obstetrics & Gynecology. 183(2):449-453, August 2000. used pure human vegf

VEGF-poly
Kitajima 2004 Gonadotropin-releasing hormone agonist administration reduced vascular endothelial growth factor (VEGF), VEGF receptors, and vascular permeability of the ovaries of hyperstimulated rats Fertility and Sterility, Volume 81, Supplement 1, March 2004, Pages 842-849 WB rat ovary

VEGFR1
Hosford GE 2003 Effects of hyperoxia on VEGF, its receptors, and HIF-2-a in the newborn rat lung Am J Physiol Lung Cell Mol Physiol 285: L161-L168, WB, rat lung
VEGFR1
Kitajima 2004 Gonadotropin-releasing hormone agonist administration reduced vascular endothelial growth factor (VEGF), VEGF receptors, and vascular permeability of the ovaries of hyperstimulated rats Fertility and Sterility, Volume 81, Supplement 1, March 2004, Pages 842-849 WB rat ovary

VEGFR1
Mayr-wohlfart U et al 2002 Vascular endothelial growth factor stimulates chemotactic migration of primary human osteoblasts Bone 30, 472-477 WB, human bone

VEGFR2
Hosford GE 2003 Effects of hyperoxia on VEGF, its receptors, and HIF-2-a in the newborn rat lung Am J Physiol Lung Cell Mol Physiol 285: L161-L168, WB, rat lung

VEGFR2
Kitajima 2004 Gonadotropin-releasing hormone agonist administration reduced vascular endothelial growth factor (VEGF), VEGF receptors, and vascular permeability of the ovaries of hyperstimulated rats Fertility and Sterility, Volume 81, Supplement 1, March 2004, Pages 842-849 WB rat ovary

VEGFR3
Ji R-C 2003 Lymphatic Network and Lymphangiogenesis in the Gastric Wall J. Histochem. Cytochem., Mar 2003; 51: 331 - 338. IHC monkey/4%PF/also SC

Notes: Antibodies usage is indicated in the following techniques: WB=Western Blot ; IHC-Immunohistochmistry; IP=Immunoprecipition; Flow=Flow cytometry;