BMP-1/Tolloid-like metalloproteases process endorepellin, the angiostatic C-terminal fragment of perlecan.
Endorepellin, the C-terminal domain of the heparan sulfate proteoglycan perlecan, possesses angiostatic activity. The terminal laminin-like globular (LG3) domain of endorepellin appears to possess most of the biological activity on endothelial cells. LG3 protein has been detected in the urine of patients with end-stage renal disease and in the amniotic fluid of pregnant women with premature rupture of fetal membranes. These findings suggest that proteolytic processing of endorepellin and the generation of LG3 might have biological significance. In this study, we have identified specific enzymes of the bone morphogenetic protein-1 (BMP-1)/Tolloid family of metalloproteases that cleave LG3 from recombinant endorepellin at the physiologically relevant site and that cleave LG3 from endogenous perlecan in cultured mouse and human cells. The BMP-1/Tolloid family of metalloproteases is thereby implicated in the processing of a major basement membrane proteoglycan and in the liberation of an anti-angiogenic factor. Using molecular modeling, site-directed mutagenesis and angiogenic assays, we further demonstrate that LG3 activity requires specific amino acids involved in Ca(2+) coordination.
Gonzalez
Eva M
EM
Department of Pathology, Anatomy and Cell Biology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Reed
Charles C
CC
Bix
Gregory
G
Fu
Jian
J
Zhang
Yue
Y
Gopalakrishnan
Bagavathi
B
Greenspan
Daniel S
DS
Iozzo
Renato V
RV
D043165:Angiostatic Proteins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D055395:Bone Morphogenetic Protein 1; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry; D002196:Capillaries / Q000254:growth & development; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology; D019812:Heparan Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008666:Metalloendopeptidases / Q000378:metabolism*; D045726:Metalloproteases / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D018919:Neovascularization, Physiologic; D010446:Peptide Fragments / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D055394:Tolloid-Like Metalloproteinases
D043165:Angiostatic Proteins; D019485:Bone Morphogenetic Proteins; D019812:Heparan Sulfate Proteoglycans; D010446:Peptide Fragments; D011506:Proteins; C498861:endorepellin protein, mouse; C071927:perlecan; D045726:Metalloproteases; D055394:Tolloid-Like Metalloproteinases; D008666:Metalloendopeptidases; C484478:TLL1 protein, human; C484477:Tll1 protein, mouse; C525705:BMP1 protein, human; C525707:Bmp1 protein, mouse; D055395:Bone Morphogenetic Protein 1; D002118:Calcium