Transglutaminase Inhibition by 2-[(2-Oxopropyl)thio]imidazolium Derivatives: Mechanism of Factor XIIIa Inactivation

Freund, Kurt; Doshi, K; Gaul, Stanley L.; Claremon, David A.; Remy, David C.; Baldwin, John J.; Pitzenberger, Steven M.; Stern, Andrew M.

doi:10.1021/bi00199a039

Cited by 108 publications

(102 citation statements)

References 72 publications

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“…The GK-21 peptide (GENPIYKSAVTTVVNPKYEGK) and the scrambled control peptide (GTAKINEPYSVTVPYGEKNKV) were chemically synthesized in tandem with the antennapedia third helix sequence (RQIKIWFQNRRMKWKK) by Peptide Protein Research. The irreversible active-site directed inhibitor R283 (1,3-dimethyl-2-[(2-oxopropyl)thio] imidazolium chloride) (4,22), was synthesized at Aston University. Cell Lines-Swiss 3T3 fibroblasts transfected with TG2 cDNA under tetracycline repressible promoter were cultured in Dulbecco's modified Eagle's medium as we previously described (23).…”

Section: Methodsmentioning

confidence: 99%

Fibronectin-Tissue Transglutaminase Matrix Rescues RGD-impaired Cell Adhesion through Syndecan-4 and β1 Integrin Co-signaling

Telci

Wang

et al. 2008

Journal of Biological Chemistry

120

165

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Heterotropic association of tissue transglutaminase (TG2) with extracellular matrix-associated fibronectin (FN) can restore the adhesion of fibroblasts when the integrin-mediated direct binding to FN is impaired using RGD-containing peptide. We demonstrate that the compensatory effect of the TG-FN complex in the presence of RGD-containing peptides is mediated by TG2 binding to the heparan sulfate chains of the syndecan-4 cell surface receptor. This binding mediates activation of protein kinase C␣ (PKC␣) and its subsequent interaction with ␤ 1 integrin since disruption of PKC␣ binding to ␤ 1 integrins with a cell-permeant competitive peptide inhibits cell adhesion and the associated actin stress fiber formation. Cell signaling by this process leads to the activation of focal adhesion kinase and ERK1/2 mitogen-activated protein kinases. Fibroblasts deficient in Raf-1 do not respond fully to the TG-FN complex unless either the full-length kinase competent Raf-1 or the kinase-inactive domain of Raf-1 is reintroduced, indicating the involvement of the Raf-1 protein in the signaling mechanism. We propose a model for a novel RGD-independent cell adhesion process that could be important during tissue injury and/or remodeling whereby TG-FN binding to syndecan-4 activates PKC␣ leading to its association with ␤ 1 integrin, reinforcement of actin-stress fiber organization, and MAPK pathway activation. Tissue transglutaminase (TG2)2 belongs to a family of enzymes that in the presence of Ca 2ϩ catalyze the post-translational modification of proteins either by covalent cross-linking or through the incorporation of primary amines. TG2 has a GTP binding/hydrolysis site that negatively modulates the Ca 2ϩ -dependent transamidating activity of the enzyme by obstructing access to the active site (1). As a consequence, the enzyme is likely to be inactive under normal Ca 2ϩ homeostasis. TG2 localizes mainly in the cytoplasm, yet recent reports also suggest its presence in the nucleus, mitochondria, at the cell surface, and in the extracellular matrix (ECM) (1-3). TG2 is translocated to the plasma membrane and was subsequently deposited into the ECM via a non-classical secretory mechanism reportedly dependent on active site conformation and on an intact N-terminal ␤-sandwich domain (4, 5) as well as on its possible association with integrins (6). Deposition of the enzyme into the ECM after cell damage and stress is important in the remodeling and/or stabilization of the several ECM proteins, such as FN (7,8). FN is particularly interesting since TG2 binds to this ECM protein with high affinity promoting wideranging effects on cell-matrix interactions, including the regulation of cell adhesion and migration, matrix assembly, and adhesion-dependent signaling (6,7,9).Cell adhesion to FN involves a series of coordinated signaling events orchestrated by numerous transmembrane receptors including the integrins and the superfamily of cell-surface proteoglycans (10, 11). The identity of the receptor-ligand pairing defines the composition of f...

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Section: Methodsmentioning

confidence: 99%

Fibronectin-Tissue Transglutaminase Matrix Rescues RGD-impaired Cell Adhesion through Syndecan-4 and β1 Integrin Co-signaling

Telci

Wang

et al. 2008

Journal of Biological Chemistry

120

165

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“…Cultures were also treated in parallel with TG2, inactivated by preincubation in 500 mM of the specific active site-directed inhibitor NTU283 (1,3-dimethyl-2[(oxopropyl)thio] imidazolium) for 6 h at 41C. 36 Any excess inhibitor was removed by dialysis against 5 mM Tris-HCl, pH 7.8, and the enzyme tested for activity prior to addition to the cell cultures. In some experiments, NTU283 or the active site-directed inhibitor NTU281 (N-benzyloxycarbonyl (CBZ)-Lphenylalanyl-6-dimethylsulfonium-5-oxo-L-norleucine) was added directly to cell cultures at 1 mM to assess the effect of endogenous transglutaminase inhibition on tube formation.…”

Section: Cell Culturementioning

confidence: 99%

Matrix changes induced by transglutaminase 2 lead to inhibition of angiogenesis and tumor growth

et al. 2005

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Administration of active TG2 to two different in vitro angiogenesis assays resulted in the accumulation of a complex extracellular matrix (ECM) leading to the suppression of endothelial tube formation without causing cell death. Matrix accumulation was accompanied by a decreased rate of ECM turnover, with increased resistance to matrix metalloproteinase-1. Intratumor injection of TG2 into mice bearing CT26 colon carcinoma tumors demonstrated a reduction in tumor growth, and in some cases tumor regression. In TG2 knockout mice, tumor progression was increased and survival rate reduced compared to wild-type mice. In wild-type mice, an increased presence of TG2 was detectable in the host tissue around the tumor. Analysis of CT26 tumors injected with TG2 revealed fibrotic-like tissue containing increased collagen, TG2-mediated crosslink and reduced organized vasculature. TG2-mediated modulation of cell behavior via changes in the ECM may provide a new approach to solid tumor therapy.

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“…To substantiate that polyamine incorporation was mediated by TG2, 1,3,dimethyl-2-[(2-oxopropyl) thio] imidazolium chloride (R283), a more specific and irreversible TG2 inhibitor, directed to the active site of the enzyme [12], was employed. Polyamine incorporation following MPP + treatment was dosedependently reduced by R283 (100 and 250 M) (Fig.…”

mentioning

confidence: 99%

The role of tissue transglutaminase in 1-methyl-4-phenylpyridinium (MPP+)-induced toxicity in differentiated human SH-SY5Y neuroblastoma cells

Beck

Girolamo

Griffin

et al. 2006

Neuroscience Letters

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Tissue transglutaminase (TG2) can induce post-translational modification of proteins, resulting in protein cross-linking or incorporation of polyamines into substrates, and can also function as a signal transducing G protein. The role of TG2 in the formation of insoluble cross-links has led to its implication in some neurodegenerative conditions. Exposure of pre-differentiated SH-SY5Y cells to the Parkinsonian neurotoxin 1-methyl-4-phenylpyridinium ion (MPP + ) resulted in significant dose-dependent reductions in TG2 protein levels, measured by probing Western blots with a TG2-specific antibody. Transglutaminase (TG) transamidating activity, on the other hand, monitored by incorporation of a polyamine pseudo-substrate into cellular proteins, was increased. Inhibitors of TG (putrescine) and TG2 (R283) exacerbated MPP + toxicity, suggesting that activation of TG2 may promote a survival response in this toxicity paradigm.

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Transglutaminase Inhibition by 2-[(2-Oxopropyl)thio]imidazolium Derivatives: Mechanism of Factor XIIIa Inactivation

Cited by 108 publications

References 72 publications

Fibronectin-Tissue Transglutaminase Matrix Rescues RGD-impaired Cell Adhesion through Syndecan-4 and β1 Integrin Co-signaling

Fibronectin-Tissue Transglutaminase Matrix Rescues RGD-impaired Cell Adhesion through Syndecan-4 and β1 Integrin Co-signaling

Matrix changes induced by transglutaminase 2 lead to inhibition of angiogenesis and tumor growth

The role of tissue transglutaminase in 1-methyl-4-phenylpyridinium (MPP+)-induced toxicity in differentiated human SH-SY5Y neuroblastoma cells

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