Print Email Facebook Twitter Proteolysis of the urokinase-type plasminogen activator receptor by metalloproteinase-12: Implication for angiogenesis in fibrin matrices Title Proteolysis of the urokinase-type plasminogen activator receptor by metalloproteinase-12: Implication for angiogenesis in fibrin matrices Author Koolwijk, P. Sidenius, N. Peters, E. Sier, C.F.M. Hanemaaijer, R. Blasi, F. van Hinsbergh, V.W.M. Gaubius Instituut TNO Publication year 2001 Abstract Pericellular proteolysis plays an important role in cell migration and the formation of new capillary structures. The plasminogen activator/plasmin and matrix degrading metalloproteinase (MMP) cascades act together in the remodeling of matrix and cell-matrix contacts. Previously we have shown that the formation of capillary structures by human foreskin microvascular endothelial cells (hMVECs) in a 3-dimensional fibrin matrix requires a functional urokinase-type plasminogen activator receptor (u-PAR). Here we report on the unexpected finding that inhibition of hMVEC-derived MMP activity by BB94 (batimastat) increased the outgrowth of capillary structures in a fibrin matrix. BB94 prevented the release of the u-PA binding domain D1 of u-PAR and thereby increased the number of functional u-PARs on hMVECs without affecting the u-PAR messenger RNA levels. Comparison of various types of protease inhibitors pointed to the prime involvement of MMP activity. Using recombinant MMPs it was shown that MMP-12 activity was able to release the D1 domain of cellularly expressed u-PAR. In addition, the expression of MMP-12 in control and basic fibroblast growth factor/tumor necrosis factor-α-stimulated hMVECs was shown by reverse transcriptase-polymerase chain reaction, suggesting that endothelial cell-derived MMP-12 may be involved in angiogenesis-related u-PAR shedding. This new mechanism of u-PAR cleavage provides new insights into the mutual interactions between the MMP and u-PA/plasmin systems. Moreover, it may be helpful in the interpretation of recent data on the use of specific MMP inhibitors in the treatment of several types of cancer. © 2001 by The American Society of Hematology. Chemicals/CAS: batimastat, 130370-60-4; Fibrin, 9001-31-4; Fibroblast Growth Factor 2, 103107-01-3; Metalloendopeptidases, EC 3.4.24.-; Phenylalanine, 63-91-2; plasminogen activator, urokinase receptors; Protease Inhibitors; Receptors, Cell Surface; Recombinant Proteins; RNA, Messenger; Thiophenes; Tumor Necrosis Factor-alpha; Urinary Plasminogen Activator, EC 188.8.131.52 Subject Binding SitesBlotting, WesternCells, CulturedEndothelium, VascularFibrinFibroblast Growth Factor 2Gene ExpressionHumansMetalloendopeptidasesMicrocirculationNeovascularization, PhysiologicPhenylalanineProtease InhibitorsReceptors, Cell SurfaceRecombinant ProteinsReverse Transcriptase Polymerase Chain ReactionRNA, MessengerThiophenesTransfectionTumor Necrosis Factor-alphaUrinary Plasminogen Activator To reference this document use: http://resolver.tudelft.nl/uuid:980027fc-3e6e-4d06-832e-e20c7d5e45a5 DOI https://doi.org/10.1182/blood.v97.10.3123 TNO identifier 236067 ISSN 0006-4971 Source Blood, 97 (10), 3123-3131 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.