Purification and characterization of a two-chain form of tissue inhibitor of metalloproteinases (TIMP) type 2 and a low molecular weight TIMP-like protein

Miyazaki, Kaoru; Funahashi, Kayano; Numata, Yasuharu; Koshikawa, Naohiko; Akaogi, Kotaro; Kikkawa, Yamato; Yasumitsu, Hidetaro; Umeda, Makoto

doi:10.1016/s0021-9258(19)85251-1

Cited by 53 publications

(7 citation statements)

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“…Proteins. TIMP-1 (35) and sAPP (16) were purified from the conditioned medium (CM) of the human bladder carcinoma cell line EJ-1, as described previously. TIMP-2free and TIMP-2-bound forms of progelatinase A were separately purified from the CM of the human glioblastoma cell line T98G, as described previously (36).…”

Section: Methodsmentioning

confidence: 99%

Novel Processing of β-Amyloid Precursor Protein Catalyzed by Membrane Type 1 Matrix Metalloproteinase Releases a Fragment Lacking the Inhibitor Domain against Gelatinase A

Higashi

Miyazaki

2003

Biochemistry

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In various mammalian cell lines, beta-amyloid precursor protein (APP) is proteolytically processed to release its NH(2)-terminal extracellular domain as a soluble APP (sAPP) that contains the inhibitor domain against gelatinase A. To investigate roles of sAPP in the regulation of gelatinase A activity, we examined the correlation between the activation of progelatinase A and processing of APP. We found that stimulation of HT1080 fibrosarcoma cells with concanavalin A led to an activation of endogenous progelatinase A and to a novel processing of APP, which releases a COOH-terminally truncated form of sAPP (sAPPtrc) into the culture medium. Reverse zymographic analysis showed that sAPPtrc lacked an inhibitory activity against gelatinase A. Analyses of production of sAPPtrc in the presence of various metalloproteinase inhibitors showed that membrane type 1 matrix metalloproteinase (MT1-MMP), an activator of progelatinase A, is most likely responsible for the production of sAPPtrc. When the concanavalin A-stimulated HT1080 cells were cultured in the condition that inhibited MT1-MMP activity, sAPP and APP were associated with the extracellular matrix deposited by the cells, whereas these gelatinase A inhibitors in the matrix were displaced by sAPPtrc after exertion of MT1-MMP activity. Taken together, these data support a model in which MT1-MMP-catalyzed release of sAPPtrc leads to reduction of the extracellular matrix-associated gelatinase A inhibitor, sAPP, thus making it feasible for gelatinase A to exert proteolytic activity only near its activator, MT1-MMP.

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Section: Methodsmentioning

confidence: 99%

Novel Processing of β-Amyloid Precursor Protein Catalyzed by Membrane Type 1 Matrix Metalloproteinase Releases a Fragment Lacking the Inhibitor Domain against Gelatinase A

Higashi

Miyazaki

2003

Biochemistry

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“…Materials. Thermolysin (protease type X from Bacillus thermoproteolyticus rokko) and cathepsin D (bovine spleen) were purchased from Sigma Chemical Co. Four matrix metalloproteinases (MMPs) were purified in proenzyme forms as reported: gelatinase A (MMP2; EC 3.4.24.24) from the human glioblastoma cell line T98G (Miyazaki et al" 1993), matrilysin (MMP9; EC 3.4.24.23) from the human rectal carcinoma cell line CaR-1 (Miyazaki et al, 1990), interstitial collagenase (MMP1; EC 3.4.24.7) from the human hepatoma cell line HLE (Umenishi et al, 1991), and stromelysin (MMP3; EC 3.4.24.17) from the rat transformed cell line RSV-BRL (Umenishi et al, 1990). Rabbit polyclonal antibodies against rat plasma Fn were raised as described (Fukai et al, 1991).…”

Section: Methodsmentioning

confidence: 99%

Release of Biological Activities from Quiescent Fibronectin by a Conformational Change and Limited Proteolysis by Matrix Metalloproteinases

Fukai

Ohtaki²,

Fujii³

et al. 1995

Biochemistry

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We reported that specific biological activities are confined to three domains of the fibronectin (Fn) molecule [Fukai et al. (1991) J. Biol. Chem. 266, 8807; Fukai et al. (1993) Biochemistry 32, 5746]: the potent ability to stimulate the adipocyte differentiation of ST-13 cells is in the amino-terminal fibrin-binding (Fib 1) domain (referred to as Fib 1 domain activity); the RGD-dependent activities that stimulate NIH-L13 cell migration and inhibit adipocyte differentiation are in the central cell-binding (Cell) domain (Cell domain activity); and the activity that stimulates cell migration in a RGD-independent manner is in the carboxyl-terminal fibrin-binding (Fib 2) domain (Fib 2 domain activity). Human plasma Fn which was purified without exposure to a denaturant, such as urea, exhibited no Fib 1, Fib 2, or Cell domain activity. By exposure to urea or surface adsorption, Fn showed Cell domain activity but not those of the Fib 1 and Fib 2 domains. Whether the cryptic domain activities are disclosed or not depended on whether or not the responsible domains were irreversibly exposed from confined environments of Fn structure as confirmed by light-scattering measurement and enzyme immunoassay using domain-specific monoclonal antibodies. We then investigated the action of matrix metalloproteinases (MMPs) in liberating the Fib 1, Cell, and Fib 2 domain activities. Matrilysin released only the Cell domain activity. In contrast, stromelysin, collagenase, and especially gelatinase A additionally liberated the Fib 1 and Fib 2 domain activities. The Fib 1, Fib 2, and Cell domains acquired much higher activities when they were freed from linkage with adjacent domains.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…Human TIMP-2-free MMP2 was purified from serum-free CM of human glioblastoma T98G cells (Miyazaki et al, 1993a). TIMP-1 and TIMP-2 were purified from serum-free CM of human bladder carcinoma EJ-1 cells (Miyazaki et al, 1993a). Hemopexin-like domain (HLD) of MMP2 was purified from human TIMP-2-free MMP2 treated with neutrophil elastase, by Reactive-Red agarose affinity chromatography as described before (Strongin et al, 1993;Rice and Banda, 1995).…”

Section: Preparation Of Mmp2 and Mmp Inhibitorsmentioning

confidence: 99%

“…The cell suspension was homogenized at 4 Њ C, nuclei removed by centrifugation at 3,000 rpm for 5 min, the supernatant was centrifuged at 15,000 rpm for 30 min, and the crude plasma membrane fraction was recovered as pellets. Immunoblotting was performed with rabbit polyclonal antibodies against human membrane type 1 MMP (MT1-MMP) and mouse mAb against human tissue inhibitor of metalloprotease-2 (TIMP-2) by reported methods (Miyazaki et al, 1993a), except that the antigen was detected by the enhanced chemiluminescence method with a NEN Life Science Products kit.…”

Section: Gelatin Zymography and Immunoblottingmentioning

confidence: 99%

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Role of Cell Surface Metalloprotease Mt1-Mmp in Epithelial Cell Migration over Laminin-5

Koshikawa

Giannelli

Cirulli

et al. 2000

The Journal of Cell Biology

593

493

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Laminin-5 (Ln-5) is an extracellular matrix substrate for cell adhesion and migration, which is found in many epithelial basement membranes. Mechanisms eliciting migration on Ln-5 need to be elucidated because of their relevance to tissue remodeling and cancer metastasis. We showed that exogenous addition of activated matrix metalloprotease (MMP) 2 stimulates migration onto Ln-5 in breast epithelial cells via cleavage of the γ2 subunit. To investigate the biological scope of this proteolytic mechanism, we tested a panel of cells, including colon and breast carcinomas, hepatomas, and immortalized hepatocytes, selected because they migrated or scattered constitutively in the presence of Ln-5. We found that constitutive migration was inhibited by BB94 or TIMPs, known inhibitors of MMPs. Limited profiling by gelatin zymography and Western blotting indicated that the ability to constitutively migrate on Ln-5 correlated with expression of plasma membrane bound MT1-MMP metalloprotease, rather than secretion of MMP2, since MMP2 was not produced by three cell lines (one breast and two colon carcinomas) that constitutively migrated on Ln-5. Moreover, migration on Ln-5 was reduced by MT1-MMP antisense oligonucleotides both in MMP2+ and MMP2− cell lines. MT1-MMP directly cleaved Ln-5, with a pattern similar to that of MMP2. The hemopexin-like domain of MMP2, which interferes with MMP2 activation, reduced Ln-5 migration in MT1-MMP+, MMP2+ cells, but not in MT1-MMP+, MMP2− cells. These results suggest a model whereby expression of MT1-MMP is the primary trigger for migration over Ln-5, whereas MMP2, which is activated by MT1-MMP, may play an ancillary role, perhaps by amplifying the MT1-MMP effects. Codistribution of MT1-MMP with Ln-5 in colon and breast cancer tissue specimens suggested a role for this mechanism in invasion. Thus, Ln-5 cleavage by MMPs may be a widespread mechanism that triggers migration in cells contacting epithelial basement membranes.

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Purification and characterization of a two-chain form of tissue inhibitor of metalloproteinases (TIMP) type 2 and a low molecular weight TIMP-like protein

Cited by 53 publications

References 0 publications

Novel Processing of β-Amyloid Precursor Protein Catalyzed by Membrane Type 1 Matrix Metalloproteinase Releases a Fragment Lacking the Inhibitor Domain against Gelatinase A

Novel Processing of β-Amyloid Precursor Protein Catalyzed by Membrane Type 1 Matrix Metalloproteinase Releases a Fragment Lacking the Inhibitor Domain against Gelatinase A

Release of Biological Activities from Quiescent Fibronectin by a Conformational Change and Limited Proteolysis by Matrix Metalloproteinases

Role of Cell Surface Metalloprotease Mt1-Mmp in Epithelial Cell Migration over Laminin-5

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