The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
H R Lijnen - One of the best experts on this subject based on the ideXlab platform.
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Stromelysin 1 mmp 3 is critical for intracranial bleeding after t pa treatment of stroke in mice
Journal of Thrombosis and Haemostasis, 2007Co-Authors: Nobuo Nagai, Y Suzuki, Kazuo Umemura, Desire Jose Collen, H R LijnenAbstract:Summary. Background: Tissue-type plasminogen activator (t-PA) is approved for treatment of ischemic stroke patients, but it may increase the risk of intracranial bleeding (ICB). Matrix metalloproteinases (MMPs), which can be activated through the plasminogen/plasmin system, may contribute to ICB after ischemic stroke. Objectives: To explore the contribution of plasminogen, MMP-3 and MMP-9 to ICB associated with t-PA treatment after ischemic stroke. Methods: Using a thrombotic middle cerebral artery occlusion (MCA-O) model, ICB was studied in mice with genetic deficiencies of plasminogen (Plg−/−), Stromelysin-1 (MMP-3−/−), or gelatinase B (MMP-9−/−) and their corresponding wild-type (WT) littermates. The induction of MMP-3 and MMP-9 was also studied in C57BL/6 WT mice. Results: ICB induced by t-PA (10 mg kg−1) was significantly less than WT in Plg−/− (P < 0.05) and MMP-3−/− (P < 0.05) but not in MMP-9−/− mice. Furthermore, administration of the broad-spectrum MMP inhibitor GM6001 after t-PA treatment reduced ICB significantly (P < 0.05) in MMP-3+/+ mice, but had no effect on MMP-3−/− mice. MMP-3 expression was significantly enhanced at the ischemic hemisphere; with placebo treatment, it was expressed only in neurons, whereas it was up-regulated in endothelial cells with t-PA treatment. Although MMP-9 expression was also significantly enhanced at the ischemic brain, the amount and the distribution were comparable in mice with and without t-PA treatment. Conclusions: Our data with gene-deficient mice thus suggest that plasminogen and MMP-3 are relatively more important than MMP-9 for the increased ICB induced by t-PA treatment of ischemic stroke.
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persistence of atherosclerotic plaque but reduced aneurysm formation in mice with Stromelysin 1 mmp 3 gene inactivation
Arteriosclerosis Thrombosis and Vascular Biology, 2001Co-Authors: J Silence, Florea Lupu, D Collen, H R LijnenAbstract:To investigate a potential role for Stromelysin-1 (MMP-3) in the development and progression of atherosclerotic lesions and aneurysm formation, mice with a deficiency of apolipoprotein E (ApoE−/−:M...
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persistence of atherosclerotic plaque but reduced aneurysm formation in mice with Stromelysin 1 mmp 3 gene inactivation
Arteriosclerosis Thrombosis and Vascular Biology, 2001Co-Authors: J Silence, Florea Lupu, D Collen, H R LijnenAbstract:To investigate a potential role for Stromelysin-1 (MMP-3) in the development and progression of atherosclerotic lesions and aneurysm formation, mice with a deficiency of apolipoprotein E (ApoE−/−:MMP-3+/+)) or with a combined deficiency of apoE and MMP-3 (ApoE−/−:MMP-3−/−) were kept on a cholesterol-rich diet for 30 weeks. Atherosclerotic lesions throughout the thoracic aorta were significantly larger in ApoE−/−:MMP-3−/− than in ApoE−/−:MMP-3+/+ mice ( P <0.05) and contained more fibrillar collagen ( P <0.01). Aneurysms in the thoracic and abdominal aortas were less frequent in ApoE−/−:MMP-3−/− than in ApoE−/−:MMP-3+/+ mice (8.5±1.7% vs 14±2.1% of sections, mean±SD, P <0.01). Immunocytochemistry revealed enhanced accumulation of macrophages in atherosclerotic lesions of ApoE−/−:MMP-3+/+ mice ( P <0.01) and expression of urokinase-type plasminogen activator (u-PA) and MMP-3 colocalizing with macrophages. Zymography confirmed the presence of u-PA and MMP-3 activity in extracts of atherosclerotic aortas. These data suggest that plasmin, generated by macrophage-secreted u-PA, activates pro-MMP-3 produced by accumulated macrophages. MMP-3 activity may then contribute to a reduction of plaque size, possibly by degradation of matrix components, and promote aneurysm formation by degradation of the elastica lamina.
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proteolytic cleavage of urokinase type plasminogen activator by Stromelysin 1 mmp 3
Biochemistry, 1998Co-Authors: F Ugwu, D Collen, B Van Hoef, A Bini, H R LijnenAbstract:Matrix metalloproteinase-3 (MMP-3, or Stromelysin-1) specifically hydrolyzes the Glu143−Leu144 peptide bond in 45-kDa single-chain urokinase-type plasminogen activator (scu-PA) and in its two-chain (tcu-PA) derivative, yielding a 17-kDa NH2-terminal domain comprising the u-PA receptor (u-PAR) binding site and a 32-kDa COOH-terminal moiety containing the serine proteinase domain of u-PA. The conversion is completely abolished in the presence of the MMP inhibitors EDTA or 1,10-phenanthroline. Biospecific interaction analysis indicates that binding of MMP-3 occurs through the 32-kDa fragment. The 32-kDa fragment derived from scu-PA (scu-PA-32k) has a specific activity of ≤500 IU/mg, but it can be activated with plasmin to a two-chain derivative (tcu-PA-32k) with a specific activity of 79 000 IU/mg. tcu-PA and tcu-PA-32k moieties derived from scu-PA-32k by plasmin or from tcu-PA by MMP-3 have comparable amidolytic activities toward the chromogenic substrate S-2444 (kcat/Km of 110 and 160 mM-1 s-1, respectivel...
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generation of an angiostatin like fragment from plasminogen by Stromelysin 1 mmp 3
Biochemistry, 1998Co-Authors: H R Lijnen, F Ugwu, And A Bini, D CollenAbstract:Matrix metalloproteinase-3 (MMP-3 or Stromelysin-1) specifically hydrolyzes the Glu59−Asn60, Pro447−Val448, and Pro544−Ser545 peptide bonds in plasminogen, yielding a 55 kDa NH2-terminal angiostatin-like domain (comprising kringles 1−4), a 14 kDa domain comprising kringle 5, and a 30 kDa domain comprising the serine proteinase domain. The conversion is completely abolished in the presence of the MMP inhibitors EDTA or 1,10-phenanthroline. Biospecific interaction analysis indicates that binding of proMMP-3 and MMP-3 to plasminogen occurs with comparable affinity (KA of 4.7 × 106 and 4.1 × 106 M-1, respectively) and is mediated via the miniplasminogen moiety (kringle 5 plus the proteinase domain) and via the catalytic domain of MMP-3. Thus, proteolytic cleavage of plasminogen by MMP-3 generates angiostatin-like fragments.
Hideaki Nagase - One of the best experts on this subject based on the ideXlab platform.
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activation of matrix metalloproteinase 9 mmp 9 via a converging plasmin Stromelysin 1 cascade enhances tumor cell invasion
Journal of Biological Chemistry, 1999Co-Authors: Noemi Ramosdesimone, Hideaki Nagase, Elizabeth Hahndantona, John Sipley, Deborah L French, James P QuigleyAbstract:Abstract Matrix metalloproteinase-9 (MMP-9) may play a critical catalytic role in tissue remodeling in vivo, but it is secreted by cells as a stable, inactive zymogen, pro-MMP-9, and requires activation for catalytic function. A number of proteolytic enzymes activate pro-MMP-9 in vitro, but the natural activator(s) of MMP-9 is unknown. To examine MMP-9 activation in a cellular setting we employed cultures of human tumor cells (MDA-MB-231 breast carcinoma cells) that were induced to produce MMP-9 over a 200-fold concentration range (0.03–8.1 nm). The levels of tissue inhibitors of metalloproteinase (TIMPs) in the induced cultures remain relatively constant at 1–4 nm. Quantitation of the zymogen/active enzyme status of MMP-9 in the MDA-MB-231 cultures indicates that even in the presence of potential activators, the molar ratio of endogenous MMP-9 to TIMP dictates whether pro-MMP-9 activation can progress. When the MMP-9/TIMP ratio exceeds 1.0, MMP-9 activation progresses, but through an interacting protease cascade involving plasmin and Stromelysin 1 (MMP-3). Plasmin, generated by the endogenous urokinase-type plasminogen activator, is not an efficient activator of pro-MMP-9, neither the secreted pro-MMP-9 nor the very low levels of pro-MMP-9 associated with intact cells. Although plasmin can proteolytically process pro-MMP-9, this limited action does not yield an enzymatically active MMP-9, nor does it cause the MMP-9 to be more susceptible to activation. Plasmin, however, is very efficient at generating active MMP-3 (Stromelysin-1) from exogenously added pro-MMP-3. The activated MMP-3 becomes a potent activator of the 92-kDa pro-MMP-9, yielding an 82-kDa species that is enzymatically active in solution and represents up to 50–75% conversion of the zymogen. The activated MMP-9 enhances the invasive phenotype of the cultured cells as their ability to both degrade extracellular matrix and transverse basement membrane is significantly increased following zymogen activation. That this enhanced tissue remodelling capability is due to the activation of MMP-9 is demonstrated through the use of a specific anti-MMP-9 blocking monoclonal antibody.
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mechanism of inhibition of the human matrix metalloproteinase Stromelysin 1 by timp 1
Nature, 1997Co-Authors: F X Gomisruth, Hideaki Nagase, Klaus Maskos, M Betz, Andreas Bergner, Robert Huber, Ko Suzuki, Naoki Yoshida, Keith Brew, Gleb BourenkovAbstract:Matrix metalloproteinases (MMPs) are zinc endopeptidases that are required for the degradation of extracellular matrix components during normal embryo development, morphogenesis and tissue remodelling 1 . Their proteolytic activities are precisely regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs) 1-5 . Disruption of this balance results in diseases such as arthritis, atherosclerosis, tumour growth and metastasis 1,2 . Here we report the crystal structure of an MMP-TIMP complex formed between the catalytic domain of human Stromelysin-1 (MMP-3) and human TIMP-1. TIMP-1, a 184-residue protein 5 , has the shape of an elongated, contiguous wedge. With its long edge, consisting of five different chain regions, it occupies the entire length of the active-site cleft of MMP-3. The central disulphide-linked segments Cys 1-Thr 2-Cys 3-Val 4 and Ser 68-Val 69 bind to either side of the catalytic zinc. Cys I bidentally coordinates this zinc, and the Thr-2 side chain extends into the large specificity pocket of MMP-3. This unusual architecture of the interface between MMP-3 and TIMP-1 suggests new possibilities for designing TIMP variants and synthetic MMP inhibitors with potential therapeutic applications.
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involvement of a region near valine 69 of tissue inhibitor of metalloproteinases timp 1 in the interaction with matrix metalloproteinase 3 Stromelysin 1
Biochemical Journal, 1997Co-Authors: Hideaki Nagase, Ko Suzuki, T E Cawston, Keith BrewAbstract:Tissue inhibitors of metalloproteinases (TIMPs) inhibit matrix metalloproteinases (MMPs) by forming a 1:1 stoichiometric complex, but the inhibition mechanism of these inhibitors is not known. Here we have investigated the reactive site of TIMP-1 by its proteinase susceptibility before and after forming a complex with MMP-3 (Stromelysin 1). When TIMP-1 was allowed to react with human neutrophil elastase, its inhibitory activity was destroyed. This resulted from cleavage of the Val 69 –Cys 70 bond. However, cleavage of this bond by neutrophil elastase was prevented when TIMP-1 formed a complex with the catalytic domain of MMP-3, and full TIMP-1 activity was restored after dissociation of the complex at pH 3.0 in the presence of EDTA. These results indicate that the region around Val 69 closely associates with an active MMP. The three-dimensional structure of the N-terminal domain of TIMP-2 elucidated by NMR studies [Williamson, Martorell, Carr, Murphy, Docherty, Freedman and Feeney (1994) Biochemistry 33 , 11745–11759] reveals that Val 69 and Cys 70 form part of an extended ridge that also includes the N-terminal section of the inhibitor. This region is probably involved in the interaction with the catalytic domains of MMPs.
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degradation of cross linked fibrin by matrix metalloproteinase 3 Stromelysin 1 hydrolysis of the gamma gly 404 ala 405 peptide bond
Biochemistry, 1996Co-Authors: Alessandra Bini, Yoshifumi Itoh, Bohdan J Kudryk, Hideaki NagaseAbstract:Abstract Matrix metalloproteinases (MMPs) can degrade a number of proteins that constitute the extracellular matrix. Previous studies have shown that atherosclerotic plaques contain substantial amounts of fibrin(ogen)-related antigen, and more recently, MMPs have been identified in such lesions. The hypothesis that MMPs play a role in the degradation of fibrinogen (Fg) and cross-linked fibrin (XL-Fb) was investigated. Fibrinogen became thrombin-unclottable when treated with matrix metalloproteinase 3 (MMP-3, Stromelysin 1) but not with matrix metalloproteinase 2 (MMP-2, gelatinase A). Incubation of XL-Fb clots (made with 125I-Fg) with MMP-3 resulted in complete lysis after 24 h. A D monomer-like fragment was generated by MMP-3 degradation of fibrinogen, XL-Fb, and fragment DD. Immunoreactivity with monoclonal antibody (MoAb)/4-2 (anti-gamma 392-406) but not with MoAb/4A5 (anti-gamma 397-411) suggested that a major cleavage site was within the sequence participating in the cross-linking of two gamma-chains. NH2-terminal sequence analysis of they gamma-chain of the D monomer-like fragment and of a dipeptide isolated from the MMP-3 digest of XL-fibrin identified the hydrolysis of the gamma Gly 404-Ala 405 peptide bond. These data indicate that the degradation of Fg and XL-Fb by MMP-3 is specific and different from plasmin. This mechanism of fibrinolysis might be of relevance in wound healing, inflammation, atherosclerosis, and other pathophysiological processes.
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immunolocalization of the matrix metalloproteinases gelatinase b and Stromelysin 1 in human endometrium throughout the menstrual cycle
Reproduction, 1996Co-Authors: Maria Jeziorska, Hideaki Nagase, Lois A. Salamonsen, David E WoolleyAbstract:Immunolocalization techniques were used to examine the distribution of the matrix metalloproteinases gelatinase B and Stromelysin 1 in human endometrial specimens, taken across the normal menstrual cycle. Gelatinase B was produced by glandular epithelial cells for approximately 7 days during the proliferative phase, with polymorphonuclear leucocytes, macrophages and eosinophils providing most of this enzyme at menstruation. There was no evidence that gelatinase B is produced by stromal cells or mast cells during the cycle. Immunoreactive gelatinase B in glandular epithelial cells was greatest during the late proliferative phase and just after ovulation; its presence in glandular secretion and the uterine fluid was optimal during the peri-implantation phase. Gelatinase B was clearly associated with an influx of polymorphonuclear leucocytes, macrophages and eosinophils just before, and during, menstruation. In contrast, immunostaining for Stromelysin 1 was much weaker than that for gelatinase B, and was present only around stromal cells and limited to microfocal locations at times coincident with stromal oedema (days 8-10 and 21-22). Both enzymes were widely distributed in specimens just before and during menstruation, and were particularly prominent in connective tissue stroma and vascular basement membranes. Specimens at the early proliferative stage were devoid of both enzymes. The data provide further evidence supporting a role for metalloproteinases in endometrial biology, not only in matrix remodelling during the cycle, but also in glandular secretions potentially relevant to blastocyst recognition and implantation. Our observations emphasize the functional importance of specific cell types and the temporal regulation of gelatinase B and Stromelysin 1 throughout the normal menstrual cycle.
Lynn M Matrisian - One of the best experts on this subject based on the ideXlab platform.
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osteopontin a novel substrate for matrix metalloproteinase 3 Stromelysin 1 and matrix metalloproteinase 7 matrilysin
Journal of Biological Chemistry, 2001Co-Authors: Renu Agnihotri, Hirotaka Haro, Howard C Crawford, Lynn M Matrisian, Matthew C Havrda, Lucy LiawAbstract:Abstract Osteopontin (OPN) is a secreted phosphoprotein shown to function in wound healing, inflammation, and tumor progression. Expression of OPN is often co-localized with members of the matrix metalloproteinase (MMP) family. We report that OPN is a novel substrate for two MMPs, MMP-3 (Stromelysin-1) and MMP-7 (matrilysin). Three cleavage sites were identified for MMP-3 in human OPN, and two of those sites were also cleaved by MMP-7. These include hydrolysis of the human Gly166-Leu167, Ala201-Tyr202 (MMP-3 only), and Asp210-Leu211 peptide bonds. Only the N-terminal Gly-Leu cleavage site is conserved in rat OPN (Gly151-Leu152). These sites are distinct from previously reported cleavage sites in OPN for the proteases thrombin or enterokinase. We found evidence for the predicted MMP cleavage fragments of OPN in vitro in tumor cell lines, and in vivo in remodeling tissues such as the postpartum uterus, where OPN and MMPs are co-expressed. Furthermore, cleavage of OPN by MMP-3 or MMP-7 potentiated the function of OPN as an adhesive and migratory stimulus in vitro through cell surface integrins. We predict that interaction of MMPs with OPN at tumor and wound healing sites in vivo may be a mechanism of regulation of OPN bioactivity.
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release of an invasion promoter e cadherin fragment by matrilysin and Stromelysin 1
Journal of Cell Science, 2001Co-Authors: Veerle Noe, Barbara Fingleton, Kathleen Jacobs, Howard C Crawford, Stefan Vermeulen, Wim F A Steelant, Erik Bruyneel, Lynn M Matrisian, Marc MareelAbstract:The function of many transmembrane molecules can be altered by cleavage and subsequent release of their ectodomains. We have investigated ectodomain cleavage of the cell-cell adhesion and signal-transducing molecule E-cadherin. The E-cadherin ectodomain is constitutively shed from the surface of MCF-7 and MDCKts.srcC12 cells in culture. Release of the 80 kDa soluble E-cadherin fragment is stimulated by phorbol-12-myristate-13-acetate and is inhibited by overexpression of the tissue inhibitor of metalloproteinases-2. The metalloproteinases matrilysin and Stromelysin-1 both cleave E-cadherin at the cell surface and release sE-CAD into the medium. The soluble E-cadherin fragment thus released inhibits E-cadherin functions in a paracrine way, as indicated by induction of invasion into collagen type I and inhibition of E-cadherin-dependent cell aggregation. Our results, therefore, suggest a novel mechanism by which metalloproteinases can influence invasion.
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tel a putative tumor suppressor modulates cell growth and cell morphology of ras transformed cells while repressing the transcription of Stromelysin 1
Molecular and Cellular Biology, 2000Co-Authors: Randy Fenrick, Howard C Crawford, Lilin Wang, John Nip, Joseph M Amann, Robert J Rooney, Jennifer Walkerdaniels, Diana L Hulboy, Michael S Kinch, Lynn M MatrisianAbstract:TEL is a member of the ETS family of transcription factors that interacts with the mSin3 and SMRT corepressors to regulate transcription. TEL is biallelically disrupted in acute leukemia, and loss of heterozygosity at the TEL locus has been observed in various cancers. Here we show that expression of TEL in Ras-transformed NIH 3T3 cells inhibits cell growth in soft agar and in normal cultures. Unexpectedly, cells expressing both Ras and TEL grew as aggregates. To begin to explain the morphology of Ras-plus TEL-expressing cells, we demonstrated that the endogenous matrix metalloproteinase Stromelysin-1 was repressed by TEL. TEL bound sequences in the Stromelysin-1 promoter and repressed the promoter in transient-expression assays, suggesting that it is a direct target for TEL-mediated regulation. Mutants of TEL that removed a binding site for the mSin3A corepressor but retained the ETS domain failed to repress Stromelysin-1. When BB-94, a matrix metalloproteinase inhibitor, was added to the culture medium of Ras-expressing cells, it caused a cell aggregation phenotype similar to that caused by TEL expression. In addition, TEL inhibited the invasiveness of Ras-transformed cells in vitro and in vivo. Our results suggest that TEL acts as a tumor suppressor, in part, by transcriptional repression of Stromelysin-1.
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coordinate expression of matrix metalloproteinase family members in the uterus of normal matrilysin deficient and Stromelysin 1 deficient mice
Endocrinology, 1997Co-Authors: Laura A Rudolphowen, John S Mudgett, Diana L Hulboy, Carole L Wilson, Lynn M MatrisianAbstract:The expression patterns of matrix metalloproteinase (MMP) family members during the murine estrous cycle and postpartum uterine involution were analyzed, and the consequence of removing specific MMPs during uterine functions was determined using mice deficient in either matrilysin (MAT) or Stromelysin-1 (STR-1). In wild-type animals, MAT, STR-1, STR-2, STR-3, and gelatinase A were consistently expressed during the most active phases of the estrous cycle, estrus and proestrus. The messenger RNA for these MMPs as well as collagenase-3 and the tissue inhibitors of metalloproteinases were also expressed during uterine involution, as determined by Northern analysis and in situ hybridization. Notably, MAT, STR-2, and collagenase-3 messenger RNA levels were elevated at early times of involution and rapidly decreased with time, whereas the transcripts for other MMPs remained elevated throughout the involution process. Involution proceeded normally in mice lacking MAT or STR-1; however, the expression of STR-1 and STR-2 was dramatically up-regulated in MAT nullizygous mice, and the expression of MAT and STR-2 was moderately up-regulated in STR-1-deficient animals. We conclude that the concerted action of several MMPs is likely to play an important role in the remodeling of the postpartum uterus, and that mechanisms that compensate for the loss of a specific MMP during this process appear to exist.
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matrix metalloproteinases are expressed during ductal and alveolar mammary morphogenesis and misregulation of Stromelysin 1 in transgenic mice induces unscheduled alveolar development
Molecular Biology of the Cell, 1995Co-Authors: J P Witty, J Wright, Lynn M MatrisianAbstract:The matrix-degrading metalloproteinases Stromelysin-1, Stromelysin-3, and gelatinase A are expressed during ductal branching morphogenesis of the murine mammary gland. Stromelysin-1 expression in particular correlates with ductal elongation, and in situ hybridization and three-dimensional reconstruction studies revealed that Stromelysin-1 mRNA was concentrated in stromal fibroblasts along the length of advancing ducts. Transgenic mice expressing an activated form of Stromelysin-1 under the control of the MMTV promoter/enhancer exhibited inappropriate alveolar development in virgin females. Ultrastructural analysis demonstrated that the basement membrane underlying epithelial and myoepithelial cells was amorphous and discontinuous compared with the highly ordered basal lamina in control mammary glands. Transgenic mammary glands had at least a twofold increase in the number of cells/unit area and a 1.4-fold increase in the percent of cycling cells by 13 wk of age compared with nontransgenic littermates. In addition, transgenic glands expressed beta-casein mRNA, but not protein, and resembled the proliferative and differentiated state of an animal between 8 and 10 days pregnant. An analysis of metalloproteinase expression in the glands of normal pregnant females demonstrated that the same matrix metalloproteinase family members, including Stromelysin-1, were expressed in connective tissue cells surrounding epithelial clusters during the time of lobuloalveolar development. These results suggest that metalloproteinases may assist in remodeling ECM during normal ductal and alveolar branching morphogenesis, and that disruption of the basement membrane by an activated metalloproteinase can affect basic cellular processes of proliferation and differentiation.
Zena Werb - One of the best experts on this subject based on the ideXlab platform.
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Stromelysin 1 regulates adipogenesis during mammary gland involution
Journal of Cell Biology, 2001Co-Authors: Caroline M Alexander, John S Mudgett, Sushma Selvarajan, Zena WerbAbstract:The matrix metalloproteinase MMP-3/Stromelysin-1 (Str1) is highly expressed during mammary gland involution induced by weaning. During involution, programmed cell death of the secretory epithelium takes place concomitant with the repopulation of the mammary fat pad with adipocytes. In this study, we have used a genetic approach to determine the role of Str1 during mammary involution. Although Str1 has been shown to induce unscheduled apoptosis when expressed ectopically during late pregnancy (Alexander, C.M., E.W. Howard, M.J. Bissell, and Z. Werb. 1996. J. Cell Biol. 135:1669–1677), we found that during post-lactational involution, mammary glands from transgenic mice that overexpress the tissue inhibitor of metalloproteinases, TIMP-1 (TO), or mice carrying a targeted mutation in Str1 showed accelerated differentiation and hypertrophy of adipocytes, while epithelial apoptosis was unaffected. These data suggest that matrix metalloproteinases (MMPs) do not induce unscheduled epithelial cell death after weaning, but instead alter the stromal microenvironment. We used adipogenic 3T3-L1 cells as a cell culture model to test the function of MMPs during adipocyte differentiation. Fibroblastic 3T3-L1 progenitor cells expressed very low levels of MMPs or TIMPs. The transcription of a number of MMP and TIMP mRNAs [Str1, MT1-MMP, (MMP-14) collagenase-3 (MMP-13), gelatinase A (MMP-2), and TIMP-1, -2 and -3] was induced in committed preadipocytes, but only differentiated adipocytes expressed an activated MMP, gelatinase A. The addition of MMP inhibitors (GM 6001 and TIMP-1) dramatically accelerated the accumulation of lipid during differentiation. We conclude that MMPs, especially Str1, determine the rate of adipocyte differentiation during involutive mammary gland remodeling.
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the matrix metalloproteinase Stromelysin 1 acts as a natural mammary tumor promoter
Oncogene, 2000Co-Authors: Mark D Sternlicht, Mina J. Bissell, Zena WerbAbstract:Extracellular matrix-degrading matrix metalloproteinases (MMPs) are invariably upregulated in epithelial cancers and are key agonists in angiogenesis, invasion and metastasis. Yet most MMPs are secreted not by the cancer cells themselves, but by stromal cells within and around the tumor mass. Because the stromal environment can influence tumor formation, and because MMPs can alter this environment, MMPs may also contribute to the initial stages of cancer development. Several recent studies in MMP-overexpressing and MMP-deficient mice support this possibility, but have required carcinogens or pre-existing oncogenic mutations to initiate tumorigenesis. Here we review the spontaneous development of premalignant and malignant lesions in the mammary glands of transgenic mice that express an autoactivating form of MMP-3/Stromelysin-1 under the control of the whey acidic protein gene promoter. These changes were absent in nontransgenic littermates and were quenched by co-expression of a human tissue inhibitor of metalloproteinases-1 (TIMP-1) transgene. Thus by altering the cellular microenvironment, Stromelysin-1 can act as a natural tumor promoter and enhance cancer susceptibility.
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impaired wound contraction in Stromelysin 1 deficient mice
Annals of Surgery, 1999Co-Authors: Kelli M Bullard, John S Mudgett, Zena Werb, Leif R Lund, Theodore N Mellin, Thomas K Hunt, Beth Ann Murphy, John Ronan, Michael J BandaAbstract:Perturbation of the balance between extracellular matrix (ECM) degradation and deposition contributes to a number of pathologic conditions characterized by abnormal healing and chronic inflammation. 1–3 Matrix metalloproteinases are expressed in migrating keratinocytes, the adjacent dermis, and granulation tissue of healing wounds, and their altered functions have been implicated in disease processes characterized by abnormal healing. 4–7 Stromelysin-1 (matrix metalloproteinase-3) degrades proteoglycans, laminin, fibronectin, the nonhelical domains of collagen types IV and IX, propeptides of type I collagen, and denatured collagens and activates collagenase-1. 8–11 It is synthesized primarily by fibroblasts and to a lesser extent by activated macrophages and keratinocytes adjacent to sites of injury. 12–14 Stromelysin-1 is found in settings where active ECM remodeling occurs, including the stroma of normally healing rabbit corneal wounds, 7 stromal cells and keratinocytes of chronic ulcers, 14 and burn wound fluid from humans. 15 With the possible exception of interstitial collagenase, metalloproteinase activities are overlapping. Compensatory mechanisms have been observed in Stromelysin-1–deficient (Str-1-/-) mice in studies of uterine involution 16 and collagen-induced arthritis. 17 We designed a study to investigate the impact of disruption of the Stromelysin-1 gene on incisional and excisional dermal wound healing in mice. In contrast to previous studies on mice with a disrupted Stromelysin-1 gene, 16,17 this study describes a phenotype in Str-1-/- mice that is not compensated by other metalloproteinases. Our results demonstrate that under the stress of excisional wound repair, deletion of Stromelysin-1 results in a failure of wound contraction and significantly delayed healing.
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the stromal proteinase mmp3 Stromelysin 1 promotes mammary carcinogenesis
Cell, 1999Co-Authors: Mark D Sternlicht, André Lochter, Carolyn J. Sympson, Mina J. Bissell, Bing Huey, Jeanphilippe Rougier, Joe W Gray, Daniel Pinkel, Zena WerbAbstract:Matrix metalloproteinases (MMPs) are invariably upregulated in the stromal compartment of epithelial cancers and appear to promote invasion and metastasis. Here we report that phenotypically normal mammary epithelial cells with tetracycline-regulated expression of MMP3/Stromelysin-1 (Str1) form epithelial glandular structures in vivo without Str1 but form invasive mesenchymal-like tumors with Str1. Once initiated, the tumors become independent of continued Str1 expression. Str1 also promotes spontaneous premalignant changes and malignant conversion in mammary glands of transgenic mice. These changes are blocked by coexpression of a TIMP1 transgene. The premalignant and malignant lesions have stereotyped genomic changes unlike those seen in other murine mammary cancer models. These data indicate that Str1 influences tumor initiation and alters neoplastic risk.
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alpha1 and alpha2 integrins mediate invasive activity of mouse mammary carcinoma cells through regulation of Stromelysin 1 expression
Molecular Biology of the Cell, 1999Co-Authors: André Lochter, Zena Werb, Marc Navre, Mina J. BissellAbstract:Tumor cell invasion relies on cell migration and extracellular matrix proteolysis. We investigated the contribution of different integrins to the invasive activity of mouse mammary carcinoma cells. Antibodies against integrin subunits α6 and β1, but not against α1 and α2, inhibited cell locomotion on a reconstituted basement membrane in two-dimensional cell migration assays, whereas antibodies against β1, but not against α6 or α2, interfered with cell adhesion to basement membrane constituents. Blocking antibodies against α1 integrins impaired only cell adhesion to type IV collagen. Antibodies against α1, α2, α6, and β1, but not α5, integrin subunits reduced invasion of a reconstituted basement membrane. Integrins α1 and α2, which contributed only marginally to motility and adhesion, regulated proteinase production. Antibodies against α1 and α2, but not α6 and β1, integrin subunits inhibited both transcription and protein expression of the matrix metalloproteinase Stromelysin-1. Inhibition of tumor cell invasion by antibodies against α1 and α2 was reversed by addition of recombinant Stromelysin-1. In contrast, Stromelysin-1 could not rescue invasion inhibited by anti-α6 antibodies. Our data indicate that α1 and α2 integrins confer invasive behavior by regulating Stromelysin-1 expression, whereas α6 integrins regulate cell motility. These results provide new insights into the specific functions of integrins during tumor cell invasion.
Donald Hupe - One of the best experts on this subject based on the ideXlab platform.
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a rationalization of the acidic ph dependence for Stromelysin 1 matrix metalloproteinase 3 catalysis and inhibition
Journal of Biological Chemistry, 2000Co-Authors: Linda L Johnson, Alexander Pavlovsky, Adam R Johnson, Jeffrey A Janowicz, Daniel F Ortwine, Claude Forsey Purchase, Andrew D White, Donald HupeAbstract:Abstract The pH dependence of matrix metalloproteinase (MMP) catalysis is described by a broad bell-shaped curve, indicating the involvement of two unspecified ionizable groups in proteolysis. Stromelysin-1 has a third pK a near 6, resulting in a uniquely sharp acidic catalytic optimum, which has recently been attributed to His224. This suggests the presence of a critical, but unidentified, S1′ substructure. Integrating biochemical characterizations of inhibitor-enzyme interactions with active site topography from corresponding crystal structures, we isolated contributions to the pH dependence of catalysis and inhibition of active site residues Glu202 and His224. The acidic pK a 5.6 is attributed to the Glu202·zinc·H2O complex, consistent with a role for the invariant active site Glu as a general base in MMP catalysis. The His224-dependent substructure is identified as a tripeptide (Pro221-Leu222-Tyr223) that forms the substrate cleft lower wall. Substrate binding induces a β-conformation in this sequence, which extends and anchors the larger β-sheet of the enzyme·substrate complex and appears to be essential for productive substrate binding. Because the PXY tripeptide is strictly conserved among MMPs, this “β-anchor” may represent a common motif required for macromolecular substrate hydrolysis. The striking acidic profile of Stromelysin-1 defined by the combined ionization of Glu202 and His224 allows the design of highly selective inhibitors.
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a rationalization of the acidic ph dependence for Stromelysin 1 matrix metalloproteinase 3 catalysis and inhibition
Journal of Biological Chemistry, 2000Co-Authors: Linda L Johnson, Alexander Pavlovsky, Adam R Johnson, Jeffrey A Janowicz, Daniel F Ortwine, Claude Forsey Purchase, Andrew D White, Chiu Fai Man, Donald HupeAbstract:The pH dependence of matrix metalloproteinase (MMP) catalysis is described by a broad bell-shaped curve, indicating the involvement of two unspecified ionizable groups in proteolysis. Stromelysin-1 has a third pK(a) near 6, resulting in a uniquely sharp acidic catalytic optimum, which has recently been attributed to His(224). This suggests the presence of a critical, but unidentified, S1' substructure. Integrating biochemical characterizations of inhibitor-enzyme interactions with active site topography from corresponding crystal structures, we isolated contributions to the pH dependence of catalysis and inhibition of active site residues Glu(202) and His(224). The acidic pK(a) 5.6 is attributed to the Glu(202).zinc.H(2)O complex, consistent with a role for the invariant active site Glu as a general base in MMP catalysis. The His(224)-dependent substructure is identified as a tripeptide (Pro(221)-Leu(222)-Tyr(223)) that forms the substrate cleft lower wall. Substrate binding induces a beta-conformation in this sequence, which extends and anchors the larger beta-sheet of the enzyme. substrate complex and appears to be essential for productive substrate binding. Because the PXY tripeptide is strictly conserved among MMPs, this "beta-anchor" may represent a common motif required for macromolecular substrate hydrolysis. The striking acidic profile of Stromelysin-1 defined by the combined ionization of Glu(202) and His(224) allows the design of highly selective inhibitors.
