The Experts below are selected from a list of 219 Experts worldwide ranked by ideXlab platform
Hideaki Nagase - One of the best experts on this subject based on the ideXlab platform.
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x ray structure of human prommp 1 new insights into Procollagenase activation and collagen binding
Journal of Biological Chemistry, 2005Co-Authors: D Jozic, Hideaki Nagase, Gleb Bourenkov, Robert Visse, Wolfram Bode, K MaskosAbstract:Vertebrate collagenases, members of the matrix metalloproteinase (MMP) family, initiate interstitial fibrillar collagen breakdown. It is essential in many biological processes, and unbalanced collagenolysis is associated with diseases such as arthritis, cancer, atherosclerosis, aneurysm, and fibrosis. These metalloproteinases are secreted from the cell as inactive precursors, Procollagenases (proMMPs). To gain insights into the structural basis of their activation mechanisms and collagen binding, we have crystallized recombinant human proMMP-1 and determined its structure to 2.2 A resolution. The catalytic metalloproteinase domain and the C-terminal hemopexin (Hpx) domain show the classical MMP-fold, but the structure has revealed new features in surface loops and domain interaction. The prodomain is formed by a three-helix bundle and gives insight into the stepwise activation mechanism of proMMP-1. The prodomain interacts with the Hpx domain, which affects the position of the Hpx domain relative to the catalytic domain. This interaction results in a "closed" configuration of proMMP-1 in contrast to the "open" configuration observed previously for the structure of active MMP-1. This is the first evidence of mobility of the Hpx domain in relation to the catalytic domain, providing an important clue toward the understanding of the collagenase-collagen interaction and subsequent collagenolysis.
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structural analysis of the alpha 2 integrin i domain Procollagenase 1 matrix metalloproteinase 1 interaction
Journal of Biological Chemistry, 2001Co-Authors: Thomas Stricker, Hideaki Nagase, Joann Dumin, S K Dickeson, L Chung, William C Parks, Samuel A SantoroAbstract:Abstract Previous studies have established that ligation of keratinocyte α2β1integrin by type I collagen induces expression of matrix metalloproteinase-1 (MMP-1) and that MMP-1 activity is required for the α2β1 integrin-dependent migration of primary keratinocytes across collagenous matrices. We now present evidence that MMP-1 binds the α2β1integrin via the I domain of the α2 integrin subunit. Using an enzyme-linked immunosorbent assay with purified human MMP-1 and recombinant α2 integrin I domain, we showed that the α2 integrin I domain specifically bound in a divalent cation-dependent manner to both the pro and active forms of MMP-1, but not to MMP-3 or MMP-13. Although both the I domain and MMP-1 bind divalent cations, MMP-1 bound, in a divalent cation-dependent manner, to α2 integrin I domains containing metal ion-dependent adhesion sites motif mutations that prevent divalent cation binding to the I domain, demonstrating that the metal ion dependence is a function of MMP-1. Using a series of MMP-1-MMP-3 and MMP-1-MMP-13 chimeras, we determined that both the linker domain and the hemopexin-like domain of MMP-1 were required for optimal binding to the I domain. The α2 integrin/MMP-1 interaction described here extends an emerging paradigm in matrix biology involving anchoring of proteinases to the cell surface to regulate their biological activities.
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Structural analysis of the alpha(2) integrin I domain/Procollagenase-1 (matrix metalloproteinase-1) interaction.
Journal of Biological Chemistry, 2001Co-Authors: Thomas Stricker, Hideaki Nagase, Joann Dumin, L Chung, William C Parks, Dickeson Sk, Samuel A SantoroAbstract:Abstract Previous studies have established that ligation of keratinocyte α2β1integrin by type I collagen induces expression of matrix metalloproteinase-1 (MMP-1) and that MMP-1 activity is required for the α2β1 integrin-dependent migration of primary keratinocytes across collagenous matrices. We now present evidence that MMP-1 binds the α2β1integrin via the I domain of the α2 integrin subunit. Using an enzyme-linked immunosorbent assay with purified human MMP-1 and recombinant α2 integrin I domain, we showed that the α2 integrin I domain specifically bound in a divalent cation-dependent manner to both the pro and active forms of MMP-1, but not to MMP-3 or MMP-13. Although both the I domain and MMP-1 bind divalent cations, MMP-1 bound, in a divalent cation-dependent manner, to α2 integrin I domains containing metal ion-dependent adhesion sites motif mutations that prevent divalent cation binding to the I domain, demonstrating that the metal ion dependence is a function of MMP-1. Using a series of MMP-1-MMP-3 and MMP-1-MMP-13 chimeras, we determined that both the linker domain and the hemopexin-like domain of MMP-1 were required for optimal binding to the I domain. The α2 integrin/MMP-1 interaction described here extends an emerging paradigm in matrix biology involving anchoring of proteinases to the cell surface to regulate their biological activities.
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hormonal regulation of collagenolysis in uterine cervical fibroblasts modulation of synthesis of Procollagenase prostromelysin and tissue inhibitor of metalloproteinases timp by progesterone and oestradiol 17 beta
Biochemical Journal, 1991Co-Authors: Takashi Sato, Y Mori, Kyoko Yamashita, Taro Hayakawa, Hideaki NagaseAbstract:Rabbit uterine cervical fibroblasts produced a large amount of matrix metalloproteinases (MMPs) such as collagenase (MMP-1) and stromelysin (MMP-3) and a small relatively amount of tissue inhibitor of metalloproteinases (TIMP). When cells were treated with progesterone or oestradiol-17 beta, both steroids concurrently decreased the level of Procollagenase and prostromelysin in the culture media and the steady-state levels of the respective mRNAs. On the other hand, the level of TIMP in the culture media and the steady-state level of its mRNA were simultaneously increased by these steroids. Similarly, the suppression of production of MMPs and the augmentation of TIMP production by both steroids were observed with interleukin 1 (IL-1)-treated cells, but the action of progesterone was more effective than that of oestradiol-17 beta in the IL-1-untreated and -treated cells. These results suggest that collagenolysis in uterine cervical fibroblasts is negatively regulated by steroid hormones via the acceleration of TIMP production and the suppression of synthesis of MMPs at the pretranslational level.
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identification of rabbit uterine cervical Procollagenase activator as rabbit matrix metalloproteinase 3 stromelysin
Comparative Biochemistry and Physiology B, 1991Co-Authors: Yoshifumi Itoh, Y Mori, K Suzuki, Takashi Sato, Hideaki NagaseAbstract:Abstract 1. 1. We previously reported an endogenous activator of Procollagenase from the culture medium of rabbit uterine cervical fibroblasts (Ishibashi et al. (1987) Biochem. J.241, 527–534). 2. 2. Similar activator was also purified and characterized from rabbit synovial fibroblasts (Vater et al. (1983) J. biol. Chem.258, 9374–9382), but its mode of activation of Procollagenase was reported to be different from that of purified activator from uterine cervical fibroblasts. 3. 3. Here we report the comparative studies of the two activators of Procollagenase and demonstrate that they are identified as matrix metalloproteinase 3 (stromelysin) by their immunological and functional criteria. The specific role of the activator in Procollagenase activation is also described.
Harald Tschesche - One of the best experts on this subject based on the ideXlab platform.
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potentiative effects of neutral proteinases in an inflamed lung relationship of neutrophil Procollagenase prommp 8 to plasmin cathepsin g and tryptase in bronchiectasis in vivo
European Respiratory Journal, 1997Co-Authors: Ruth Sepper, Harald Tschesche, Timo Sorsa, Y T Konttinen, Kari K Eklund, Anneli Lauhio, A O Aasen, H SillastuAbstract:We attempted to study the possible relationships between neutrophil-type Procollagenase/pro-matrix metalloproteinase (MMP-8) and the serine proteinases plasmin, cathepsin G and tryptase in bronchiectasis. The presence of the plasmin/plasminogen system and plasmin-, cathepsin G- and tryptase-like activities were compared to the activity of endogenously activated MMP-8 in bronchoalveolar lavage (BAL) fluid in 38 bronchiectasis patients and in 14 healthy controls by means of immunohistochemistry, Western-blot and substrate-based functional assays. In contrast to cathepsin G- and tryptase-like activities, the plasmin/plasminogen activator system in BAL fluid was observed to have a relatively weak activation stage and no correlation with disease severity. Neither plasmin-like activities nor concentrations of plasminogen activators from the bronchiectatic patients differed significantly from the values of healthy controls. Immunolocation of plasminogen activator inhibitor-1 showed a marked, but not significant, increase in bronchiectatic lung as compared to controls. In contrast to cathepsin G- and tryptase-like activities, with their strong and significant correlation with endogenously activated collagenase (r=0.9; p=0.0001; and r=0.6; p=0.03, respectively), no correlations were observed between plasmin-like and endogenously activated collagenase (r=0.3; p=0.2) in bronchiectasis. These findings suggest that cathepsin G- and tryptase-like activities may act as potent pro-matrix metalloproteinase-8 activators in patients with bronchiectasis, whereas the plasminogen activator/plasmin cascade was shown to be down-regulated.
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activation of human neutrophil Procollagenase by stromelysin 2
FEBS Journal, 1996Co-Authors: Vera Knäuper, Gillian Murphy, Harald TschescheAbstract:Neutrophil Procollagenase (MMP-8) was efficiently activated by incubation with active stromelysin 2 (MMP-10). A single-step activation mechanism involving the cleavage of the Gly78-Phe79 peptide bond at the end of the propeptide domain was observed. Determination of the collagenolytic activity revealed the generation of active neutrophil collagenase displaying high specific activity. When compared with the specific activity following mercurial activation, which generates active collagenase by autoproteolytic cleavage of either Phe79-Met80 or Met80-Leu81 peptide bonds [Blaser, J., Knauper, V., Osthues, A., Reinke, H. & Tschesche, H. (1991) Eur J. Biochem. 202, 1223–1230], the specific activity of the stromelysin-2-activated enzyme was considerably higher. Thus, human neutrophil Procollagenase was ‘superactivated’ by stromelysin 2, as was recently shown for the stromelysin-1-activated enzyme [Knauper, V., Wilhelm, S. M., Seperack, P. K., De Clerck, Y. A., Langley, K. E., Osthues, A. & Tschesche, H. (1993a) Biochem. J. 295, 581–586].
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direct activation of human neutrophil Procollagenase by recombinant stromelysin
Biochemical Journal, 1993Co-Authors: V Knäuper, Yves A Declerck, Keith Langley, Anja Osthues, S Wilhelm, P K Seperack, Harald TschescheAbstract:Human neutrophil Procollagenase was activated by incubation with recombinant active stromelysin. Activation was achieved by cleavage of the Gly78-Phe79 peptide bond at the end of the propeptide domain in a single-step activation mechanism. In addition, accelerated activation was achieved when N-terminally truncated, latent collagenase (with Phe49 as its N-terminal residue) was incubated with recombinant active stromelysin. Determination of the specific activity of recombinant-stromelysin-activated neutrophil collagenase with dinitrophenyl-octapeptide or type I collagen demonstrated the generation of high specific activity. The specific activity of stromelysin-activated enzyme was considerably higher than that of trypsin- or HgCl2-activated collagenase. Thus human neutrophil collagenase is superactivated, like the homologous fibroblast collagenase [Murphy, Cockett, Stephens, Smith and Docherty (1987) Biochem. J. 248, 265-268]. The occurrence of Phe79 at the N-terminus of the neutrophil collagenase seemed to be critical for superactivation, which is in agreement with data published by Suzuki, Enghild, Morodomi, Salvesen and Nagase [(1990) Biochemistry 29, 10261-10270] on fibroblast collagenase.
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The recombinant catalytic domain of human neutrophil collagenase lacks type I collagen substrate specificity.
Biochemical and Biophysical Research Communications, 1993Co-Authors: S. Schnierer, T. Kleine, T. Gote, A. Hillemann, V Knäuper, Harald TschescheAbstract:Abstract The coding region for human neutrophil short form Procollagenase lacking the hemopexin like domain coding region was amplified by polymerase chain reaction. Recombinant short form Procollagenase was expressed in E.coli and purified in a three step procedure. Renaturation of this proenzyme was carried out by an effective new method using Q-Sepharose chromatography. Treatment of short form Procollagenase with mercurials resulted in active short form collagenase M r 21000 and an intermediate product of M r 23000. These two products were separated by hydroxamate affinity chromatography. The active, short form collagenase M r 21000 is stable. Despite full proteolytic activity, it lacks type I collagen substrate specificity and forms the basis for crystallisation experiments.
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formation of a covalent hg cys bond during mercurial activation of pmnl Procollagenase gives evidence of a cysteine switch mechanism
FEBS Letters, 1992Co-Authors: J Blaser, Heinz Reinke, Susanne Triebel, Harald TschescheAbstract:A common method for the activation of mammalian matalloproteinases is the use of mercurial compounds. Activation of PMNL Procollagenase by soluble mercurials takes place as a three-step mechanism with a final intermolecular loss of the PRCGVPD autoinhibitor region. In this study covalently bound mercury in the form of mercurial agarose was chosen to probe activation of PMNL Procollagenase. Activation was not achieved, since the final intermolecular cleavage with removal of the PRCGVPD motif could not take place. An intermediate form of the enzyme was bound to the column. Its N-terminal sequence determination proved cleavage of the Asp64-Met65 peptide bond leaving the cysteine of the propeptide domain for covalent attachment to the mercurial agarose. This gives further evidence of a cysteine-switch mechanism involving Cys71.
William G Stetlerstevenson - One of the best experts on this subject based on the ideXlab platform.
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cellular activation of the 72 kda type iv Procollagenase timp 2 complex
Kidney International, 1993Co-Authors: Peter D Brown, David E Kleiner, Edward J Unsworth, William G StetlerstevensonAbstract:Cellular activation of the 72 kDa type IV Procollagenase/TIMP-2 complex. Members of the collagenase family of enzymes have been implicated as central mediators of a number of both physiologic and pathologic processes. The 72-kDa type IV collagenase is secreted as a latent proenzyme, complexed with tissue inhibitor of metalloprotein-ase-2 (TIMP-2). Like other members of the collagenase family, this enzyme complex must be converted to a catalytically active form for proteolytic remodeling of extracellular matrix to occur. In the current study we demonstrate an inducible cell-mediated activation of the 72-kDa type IV Procollagenase/TIMP-2 complex. Isolation of the 62 kDa activated enzyme/TIMP-2 complex from conditioned media of concanavalin A treated WI-38 fibroblasts demonstrated that the cell activated species was proteolytically active and amino terminal sequencing gave the sequence YNFF. This is identical to that of the 62 kDa species generated following organomercurial activation of purified 72-kDa type IV Procollagenase/TIMP-2 complex. We have also isolated biosynthetically 35 S-labeled 72-kDa type IV Procollagenase/TIMP-2 complex and used this to further study the cellular activation process. In cell lines tested the activator was retained in the residual cell fraction following lysis in the presence of 0.2% (wt/vol) Brij-35. Inhibitor studies demonstrated that processing and activation of 72-kDa type IV Procollagenase/TIMP-2 complex by the residual fraction was inhibited by 5 mM ethylenediaminetetraacetic acid and 0.5 mM 1,10-phenanthroline demonstrating a metal atom dependence. The species responsible for activation could be partially recovered in soluble form with 0.5% (vol/vol) Triton X-100 and 0.25% (wt/vol) CHAPS but was not salt extractable. This cellular activation process differs from that demonstrated for interstitial collagenase in that it is not mediated by soluble factors and is not plasminogen activator/plasmin dependent.
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secretion of latent type iv Procollagenase and active type iv collagenase by testicular cells in culture
Biochemical Journal, 1991Co-Authors: Menachem Ailenberg, William G Stetlerstevenson, I B FritzAbstract:Testicular peritubular myoid cells, which have properties similar to those of vascular smooth-muscle cells, secrete a variety of metalloproteinases when maintained in culture in a chemically defined medium. The predominant metalloproteinases secreted were identified as latent type IV Procollagenases having molecular masses of 72 kDa and 75 kDa, as detected in Western immunoblots with specific antibodies against type IV Procollagenase. When peritubular cells were stimulated by dibutyryl cyclic AMP, forskolin or cholera toxin, they secreted increased amounts of type IV Procollagenase. However, little if any of the active type IV collagenase, having a lower molecular mass of 66 kDa, could be detected under these conditions. Addition of low concentrations of cytochalasin D to peritubular cells in monoculture resulted in conversion of the latent type IV collagenase into its active form, assessed with antibody-specificity studies and by the appearance of the 66 kDa protein. In contrast, Sertoli cells in culture did not manifest an increased conversion of type IV Procollagenase into type IV collagenase in the presence of cytochalasin D, even though cytochalasin D addition invariably resulted in a disruption of the microfilament assembly in each of these gonadal somatic cell populations. When peritubular cells were co-cultured with Sertoli cells, addition of cytochalasin D no longer resulted in formation of increased amounts of the active form of type IV collagenase. Sertoli cells and peritubular cells each secreted a tissue inhibitor of metalloproteinase type 2, detected with a specific antibody in a Western immunoblot to have a molecular mass of 21 kDa. We conclude that cytochalasin D acts on mesenchymal-type peritubular cells, but not on epithelial-type Sertoli cells, to enhance the conversion of latent type IV Procollagenase into active type IV collagenase. This conversion of type IV Procollagenase into type IV collagenase by peritubular cells was inhibited by factor(s) secreted by Sertoli cells. Interactions between Sertoli cells and peritubular cells are postulated to modulate net proteinase activities in discrete regions of the testis.
Y Mori - One of the best experts on this subject based on the ideXlab platform.
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Urinary trypsin inhibitor suppresses the production of interstitial Procollagenase/proMMP-1 and prostromelysin 1/proMMP-3 in human uterine cervical fibroblasts and chorionic cells
FEBS Letters, 1997Co-Authors: Keisuke Imada, Naohiro Kanayama, Toshihiko Terao, Y MoriAbstract:The mechanisms by which urinary trypsin inhibitor (UTI) prevents preterm premature rupture of fetal membrane and premature cervical ripening were investigated. We, therefore, examined the effects of UTI on the production of matrix metalloproteinases (MMPs) which closely participate in the breakdown of extracellular matrix in cultured human uterine cervical fibroblasts and human chorionic cells. UTI suppressed specifically the production of interstitial Procollagenase/proMMP-1 and prostromelysin 1/proMMP-3 from both cells in a dose-dependent manner (0.32-1.28 μM). This suppression was accompanied by a decrease in steady-state levels of their mRNAs. These results indicate for the first time that UTI down-regulates the production of proMMP-1 and proMMP-3 accompanying with the decrease in the expression of their mRNAs, and therefore UTI actually participates in the maintenance of fetal membranes and/or uterine cervix by overall suppression of MMP production along with the known inhibitory actions towards serine proteinases.
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urinary trypsin inhibitor suppresses the production of interstitial Procollagenase prommp 1 and prostromelysin 1 prommp 3 in human uterine cervical fibroblasts and chorionic cells
FEBS Letters, 1997Co-Authors: Keisuke Imada, Naohiro Kanayama, Toshihiko Terao, Y MoriAbstract:The mechanisms by which urinary trypsin inhibitor (UTI) prevents preterm premature rupture of fetal membrane and premature cervical ripening were investigated. We, therefore, examined the effects of UTI on the production of matrix metalloproteinases (MMPs) which closely participate in the breakdown of extracellular matrix in cultured human uterine cervical fibroblasts and human chorionic cells. UTI suppressed specifically the production of interstitial Procollagenase/proMMP-1 and prostromelysin 1/proMMP-3 from both cells in a dose-dependent manner (0.32-1.28 μM). This suppression was accompanied by a decrease in steady-state levels of their mRNAs. These results indicate for the first time that UTI down-regulates the production of proMMP-1 and proMMP-3 accompanying with the decrease in the expression of their mRNAs, and therefore UTI actually participates in the maintenance of fetal membranes and/or uterine cervix by overall suppression of MMP production along with the known inhibitory actions towards serine proteinases.
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hormonal regulation of collagenolysis in uterine cervical fibroblasts modulation of synthesis of Procollagenase prostromelysin and tissue inhibitor of metalloproteinases timp by progesterone and oestradiol 17 beta
Biochemical Journal, 1991Co-Authors: Takashi Sato, Y Mori, Kyoko Yamashita, Taro Hayakawa, Hideaki NagaseAbstract:Rabbit uterine cervical fibroblasts produced a large amount of matrix metalloproteinases (MMPs) such as collagenase (MMP-1) and stromelysin (MMP-3) and a small relatively amount of tissue inhibitor of metalloproteinases (TIMP). When cells were treated with progesterone or oestradiol-17 beta, both steroids concurrently decreased the level of Procollagenase and prostromelysin in the culture media and the steady-state levels of the respective mRNAs. On the other hand, the level of TIMP in the culture media and the steady-state level of its mRNA were simultaneously increased by these steroids. Similarly, the suppression of production of MMPs and the augmentation of TIMP production by both steroids were observed with interleukin 1 (IL-1)-treated cells, but the action of progesterone was more effective than that of oestradiol-17 beta in the IL-1-untreated and -treated cells. These results suggest that collagenolysis in uterine cervical fibroblasts is negatively regulated by steroid hormones via the acceleration of TIMP production and the suppression of synthesis of MMPs at the pretranslational level.
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identification of rabbit uterine cervical Procollagenase activator as rabbit matrix metalloproteinase 3 stromelysin
Comparative Biochemistry and Physiology B, 1991Co-Authors: Yoshifumi Itoh, Y Mori, K Suzuki, Takashi Sato, Hideaki NagaseAbstract:Abstract 1. 1. We previously reported an endogenous activator of Procollagenase from the culture medium of rabbit uterine cervical fibroblasts (Ishibashi et al. (1987) Biochem. J.241, 527–534). 2. 2. Similar activator was also purified and characterized from rabbit synovial fibroblasts (Vater et al. (1983) J. biol. Chem.258, 9374–9382), but its mode of activation of Procollagenase was reported to be different from that of purified activator from uterine cervical fibroblasts. 3. 3. Here we report the comparative studies of the two activators of Procollagenase and demonstrate that they are identified as matrix metalloproteinase 3 (stromelysin) by their immunological and functional criteria. The specific role of the activator in Procollagenase activation is also described.
Vera Knäuper - One of the best experts on this subject based on the ideXlab platform.
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Investigation of the role of Endo180/urokinase-type plasminogen activator receptor-associated protein as a collagenase 3 (matrix metalloproteinase 13) receptor
Biochemical Journal, 2002Co-Authors: Louise Bailey, Dirk Wienke, Matthew J. Howard, Vera Knäuper, Clare M. Isacke, Gillian MurphyAbstract:Procollagenase 3 can be activated by interaction with and cleavage by the cell-associated membrane type 1 metalloproteinase (MT1 MMP; MMP 14). It has also been shown to bind to a specific receptor, and is subsequently internalized via the low-density lipoprotein-related receptor by osteoblast cell lines. The receptor was identified as a recycling glycoprotein of the macrophage mannose receptor family, Endo180. In order to ascertain whether there is a relationship between Endo180 binding and Procollagenase 3 activation, we have compared Procollagenase 3 activation by an HT1080 fibrosarcoma cell line overexpressing MT1 MMP, without and with overexpression of Endo180. No difference in Procollagenase 3 activation was observed, and neither was the enzyme bound to the cells or internalized. In contrast, the osteoblast cell lines, MG63 and UMR-106, both bound and internalized Procollagenase 3. However, immunolocalization studies showed that the Endo180 abundantly expressed by these cells did not co-localize with the Procollagenase 3. In further biochemical studies we confirmed that Procollagenase 3 did not bind to Endo180, using both ligand- blotting and immunoprecipitation techniques. We conclude that Endo180 is unlikely to be a receptor for collagenase 3 in relation to either its activation or cell binding and internalization, and that other interaction partners must be sought.
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the role of the c terminal domain of human collagenase 3 mmp 13 in the activation of Procollagenase 3 substrate specificity and tissue inhibitor of metalloproteinase interaction
Journal of Biological Chemistry, 1997Co-Authors: Vera Knäuper, Carlos Lopezotin, Bryan J Smith, Susan Cowell, Mark Oshea, Helen Morris, Luciano Zardi, Gillian MurphyAbstract:Abstract Recombinant human Procollagenase-3 and a C-terminal truncated form (Δ249-451 Procollagenase-3) have been stably expressed in myeloma cells and purified. The truncated proenzyme could be processed by aminophenylmercuric acetate via a short-lived intermediate form (N-terminal Leu58) to the final active form (N-terminal Tyr85). The kinetics of activation were not affected by removal of the hemopexin-like C-terminal domain. The specific activities of both collagenase-3 and Δ249-451 collagenase-3 were found to be similar using two quenched fluorescent substrates, but Δ249-451 collagenase-3 failed to cleave native triple helical collagens (types I and II) into characteristic one- and three-quarter fragments. It was noted, however, that the β1,2(I) chains of type I collagen were susceptible to Δ249-451 collagenase-3, which indicates that the catalytic domain displays telopeptidase activity, thereby generating α1,2(I) chains that are slightly shorter than those in native type I collagen. It can be concluded that the C-terminal domain is only essential for the triple helicase activity of collagenase-3. Binding of Procollagenase-3 and active collagenase-3 to type I collagen is mediated by the C-terminal domain. Both collagenase-3 and Δ249-451 collagenase-3 hydrolyzed the large tenascin C isoform, fibronectin, recombinant fibronectin fragments, and type IV, IX, X, and XIV collagens; thus, these events were independent from C-terminal domain interactions. In contrast, the minor cartilage type XI collagen was resistant to cleavage. Kinetic analysis of the mechanism of inhibition of wild-type and Δ249-451 collagenase-3 by wild-type and mutant tissue inhibitors of metalloproteinase (TIMPs) revealed that the association rates for complex formation were influenced by both N- and C-terminal domain interactions. The C-terminal domain of wild-type collagenase-3 promoted increased association rates with the full-length inhibitors TIMP-1 and TIMP-3 and the hybrid N.TIMP-2/C.TIMP-1 by a factor of up to 33. In contrast, the association rates for complex formation with TIMP-2 and N.TIMP-1/C.TIMP-2 were only marginally affected by C-terminal domain interactions.
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cellular mechanisms for human Procollagenase 3 mmp 13 activation evidence that mt1 mmp mmp 14 and gelatinase a mmp 2 are able to generate active enzyme
Journal of Biological Chemistry, 1996Co-Authors: Vera Knäuper, Horst Will, Carlos Lopezotin, Bryan J Smith, Susan J Atkinson, Heather Stanton, R M Hembry, Gillian MurphyAbstract:Abstract Gelatinase A and membrane-type metalloproteinase (MT1-MMP) were able to process human Procollagenase-3 (Mr 60,000) to the fully active enzyme (Tyr85 N terminus; Mr 48,000). MT1-MMP activated Procollagenase-3 via a Mr 56,000 intermediate (Ile36 N terminus) to 48,000 which was the result of the cleavage of the Glu84-Tyr85 peptide bond. We have established that the activation rate of Procollagenase-3 by MT1-MMP was enhanced in the presence of progelatinase A, thereby demonstrating a unique new activation cascade consisting of three members of the matrix metalloproteinase family. In addition, Procollagenase-3 can be activated by plasmin, which cleaved the Lys38-Glu39 and Arg76-Cys77 peptide bonds in the propeptide domain. Autoproteolysis then resulted in the release of the rest of the propeptide domain generating Tyr85 N-terminal active collagenase-3. However, plasmin cleaved the C-terminal domain of collagenase-3 which results in the loss of its collagenolytic activity. Concanavalin A-stimulated fibroblasts expressing MT1-MMP and fibroblast-derived plasma membranes were able to process human Procollagenase-3 via a Mr 56,000 intermediate form to the final Mr 48,000 active enzyme which, by analogy with progelatinase A activation, may represent a model system for in vivo activation. Inhibition experiments using tissue inhibitor of metalloproteinases, plasminogen activator inhibitor-2, or aprotinin demonstrated that activation in the cellular model system was due to MT1-MMP/gelatinase A and excluded the participation of serine proteinases such as plasmin during Procollagenase-3 activation. We have established that progelatinase A can considerably potentiate the activation rate of Procollagenase-3 by crude plasma membrane preparations from concanavalin A-stimulated fibroblasts, thus confirming our results using purified progelatinase A and MT1-MMP. This new activation cascade may be significant in human breast cancer pathology, where all three enzymes have been implicated as playing important roles.
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activation of human neutrophil Procollagenase by stromelysin 2
FEBS Journal, 1996Co-Authors: Vera Knäuper, Gillian Murphy, Harald TschescheAbstract:Neutrophil Procollagenase (MMP-8) was efficiently activated by incubation with active stromelysin 2 (MMP-10). A single-step activation mechanism involving the cleavage of the Gly78-Phe79 peptide bond at the end of the propeptide domain was observed. Determination of the collagenolytic activity revealed the generation of active neutrophil collagenase displaying high specific activity. When compared with the specific activity following mercurial activation, which generates active collagenase by autoproteolytic cleavage of either Phe79-Met80 or Met80-Leu81 peptide bonds [Blaser, J., Knauper, V., Osthues, A., Reinke, H. & Tschesche, H. (1991) Eur J. Biochem. 202, 1223–1230], the specific activity of the stromelysin-2-activated enzyme was considerably higher. Thus, human neutrophil Procollagenase was ‘superactivated’ by stromelysin 2, as was recently shown for the stromelysin-1-activated enzyme [Knauper, V., Wilhelm, S. M., Seperack, P. K., De Clerck, Y. A., Langley, K. E., Osthues, A. & Tschesche, H. (1993a) Biochem. J. 295, 581–586].
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mercurial activation of human pmn leucocyte type iv Procollagenase gelatinase
FEBS Letters, 1992Co-Authors: Susanne Triebel, Vera Knäuper, Heinz Reinke, J Blaser, Harald TschescheAbstract:Autoproteolytic activation and processing of human polymorphonuclear leucocyte (PMNL) type IV Procollagenase (gelatinase) was initiated by HgCl2 and was investigated by kinetic analysis and N-terminal sequence determination of the reaction products. In the first instance the propeptide domain was lost by subsequent cleavage of the Asp15-Leu16, Glu40-Met41, Leu52-Leu53 and Ala74-Met75 peptide bonds. The PRCGVPD sequence motif (residues Pro78-Asp84), which is conserved in all metalloproteinases and expected to be relevant for latency, remained uncleaved at the activated enzyme. The generated intermediate was further processed by three C-terminal cleavages. The Glu666-Leu667, Ala506-Glu507 and Ala398-Leu399 bonds were hydrolysed sucessively. From the fragmentation products we were able to conclude that three released fragment peptides contained unpaired free cysteine with the residues Cys497, Cys653, Cys683. Cleavage of the first C-terminal peptide bond resulted in the loss of one of the conserved Cys residues of the hemopexin-like domain, whereas the Cys residue of the PRCGVPD motif was retained at the fully active enzyme. The possibility of an entirely different activation mechanism for PMNL type IV Procollagenase is discussed.