Transamidase kinetics. Amide formation in the enzymic reactions of thiol esters with amines

Stenberg, P.; Curtis, C. G.; Wing, Deborah A.; Tong, Yam Shun; Credo, R.B.; Gray, A.; Lóránd, L.

doi:10.1042/bj1470153

Cited by 45 publications

(21 citation statements)

References 24 publications

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“…Although there are substantial similarities between the kinetic pathways of TGs and the evolutionarily related papain family of enzymes in that both proceed by consecutive steps of acylation and deacylation, a number of major differences are also evident (21)(22)(23)(24). Just as papain, TGs can operate in a hydrolytic mode and, just as the TGs, the papain family of proteases can also function well in transamidating and transesterification reactions.…”

Section: Resultsmentioning

confidence: 99%

Conserved tryptophan in the core domain of transglutaminase is essential for catalytic activity

Murthy

Iismaa

Begg

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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T ransglutaminase 2 [here designated TG2 (EC2.3.2.13), but also identified as the GTP-binding protein, G h ] differs from its relatives in the family of Ca 2ϩ -dependent protein crosslinking enzymes mainly by its affinity for nucleotides (1-3) and for fibronectin (4-8). GTP-binding and hydrolysis enable TG2 to act as a G protein in signal transduction (G h ; ref. 9); GTP also serves as a potent allosteric inhibitor that suppresses the Ca 2ϩ -activated crosslinking activities of the enzyme. Independently of any of its catalytic functions and GTP-binding activity, TG2 binds fibronectin with very high affinity (7). Thus, when expressed on cell surfaces, TG2 plays an important role in organizing the extracellular matrix as an integrin-binding adhesion coreceptor (10, 11).Crystal structures are available for two related proteins: the A subunit of the coagulation factor XIII zymogen (fXIIIA; ref. 12) and the sea bream liver TG (13). Although neither of these is regulated by GTP (personal information from Kohki Ishikawa regarding the latter enzyme), sequence homologies between TG2 (rat) and these proteins suggest that TG2͞G h also would be organized into four domains: a ␤ sandwich (residues 1-138) followed by a large ␣͞␤ catalytic core (139-471) and two ␤ barrels [barrel 1 (472-584) and barrel 2 (585-686)]. The N-terminal (Ϸ28 kDa) region of TG2 is responsible for binding fibronectin (8) whereas the C-terminal segment of barrel 2 could interact with phospholipase in signal transduction (14). The core domain comprises the papain-like catalytic center (15) and the nucleotidebinding residues (16). Like the monomeric but unlike the heterotrimeric G proteins, TG2 was shown to bind 2Ј-(or 3Ј)-O-(Nmethylanthraniloyl)GTP (mantGTP) with high affinity (17). Binding was evidenced also by the appearance of an energy transfer band from one or more Trp residues of the protein to the mant fluorophore in this nucleotide. Because the core domain contains almost all of the tryptophans conserved throughout the TG family, we evaluated the nucleotide-binding properties and the transamidating activities of candidate tryptophan point mutants of TG2. Specifically, W180, 241, 278, 332, and 337 were targeted because they surround S171, a residue identified to be critical for GTPbinding. On a model of TG2 based on the fXIIIA zymogen crystal structure (16), all of the mutated residues are within 20Å (range 9.3-18.7Å) of both S171 and the active-site cysteine C277. We show that no single Trp residue alone accounts for the observed energy transfer from the protein to the nucleotide but some of the tryptophan mutants displayed greatly diminished mantGTPbinding affinities. Another important finding was that residue W241-that is highly conserved among the TGs but is lacking in the catalytically inactive family member of human red cell, band 4.2-is critical for transamidation by TG2. Materials and MethodsConstructs. Site-directed mutants of rat TG2͞G h cDNA were generated in a temperature-cycling reaction with Pfu DNA polymerase by using two complement...

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Section: Resultsmentioning

confidence: 99%

Conserved tryptophan in the core domain of transglutaminase is essential for catalytic activity

Murthy

Iismaa

Begg

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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“…RA-induced Activation of RhoA Is Mediated by TGase-To determine the role of TGase in activation of RhoA, we have used monodansylcadaverine (MDC), a known TGase inhibitor (45), and as shown in Fig. 3A, MDC dose-dependently inhibited RA-induced transglutaminase activity.…”

Section: Ra Promotes Expression Of Tgase and Neuronal Markers-mentioning

confidence: 99%

Tissue Transglutaminase Mediates Activation of RhoA and MAP Kinase Pathways during Retinoic Acid-induced Neuronal Differentiation of SH-SY5Y Cells

Singh¹,

Pan²,

Kao

et al. 2003

Journal of Biological Chemistry

146

153

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All-trans-retinoic acid (RA) plays a crucial role in survival and differentiation of neurons. For elucidating signaling mechanisms involved in RA-induced neuronal differentiation, we have selected SH-SY5Y cells, which are an established in vitro cell model for studying RA signaling. Here we report that RA-induced neuronal differentiation of SH-SY5Y cells is coupled with increased expression/activation of TGase and in vivo transamidation and activation of RhoA. In addition, RA promotes formation of stress fibers and focal adhesion complexes, and activation of ERK1/2, JNK1, and p38␣/␤/␥ MAP kinases. Using C-3 exoenzyme (RhoA inhibitor) or monodansylcadaverine (TGase inhibitor), we show that transamidated RhoA regulates cytoskeletal rearrangement and activation of ERK1/2 and p38␥ MAP kinases. Further, by using stable SH-SY5Y cell lines (overexpressing wild-type, C277S mutant, and antisense TGase), we demonstrate that transglutaminase activity is required for activation of RhoA, ERK1/2, JNK1, and p38␥ MAP kinases. Activated MAP kinases differentially regulate RA-induced neurite outgrowth and neuronal marker expression. The results of our studies suggest a novel mechanism of RA signaling, which involves activation of TGase and transamidation of RhoA. RA-induced activation of TGase is proposed to induce multiple signaling pathways that regulate neuronal differentiation.

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“…Another influence on TG activity could be the presence of amine compounds that are known to competitively inhibit the enzyme (Ishitani and Suzuki, 1989;Martinez et al, 1989;Stenberg et al, 1975). Degranulating mast cells, which release histamine, an inhibitor of TG (Martinez et al, 1989), are a component of the cellular inflammatory response in the plaque (Kaartinen et al, 1998;Kovanen et al, 1995;Lendon et al, 1991, van der Wal et al, 1994.…”

Section: Tg In Atherosclerosismentioning

confidence: 99%

Localization of Tissue Transglutaminase in Human Carotid and Coronary Artery Atherosclerosis: Implications for Plaque Stability and Progression

Haroon

Wannenburg

Gupta

et al. 2001

Laboratory Investigation

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SUMMARY:Although atherosclerosis progresses in an indolent state for decades, the rupture of plaques creates acute ischemic syndromes that may culminate in myocardial infarction and stroke. Mechanical forces and matrix metalloproteinase activity initiate plaque rupture, whereas tissue inhibitors of metalloproteinases have an important (albeit indirect) role in plaque stabilization. In this paper, an enzyme that could directly stabilize the plaque is described. Tissue transglutaminase (TG) catalyzes the formation of ⑀(␥-glutamyl)lysine isopeptide bonds that are resistant to enzymatic, mechanical, and chemical degradation. We performed immunohistochemistry for TG in atherosclerotic human coronary and carotid arteries. TG was most prominent along the luminal endothelium and in the medium of the vessels with a distribution mirroring that of smooth muscle cells. Variable, often prominent, immunoreactivity for TG was also seen in the intima, especially in regions with significant neovascularization. Additionally, TG was detected in fibrous caps and near the "shoulder regions" of some plaques. A monoclonal antibody to the transglutaminase product ⑀(␥-glutamyl)lysine isopeptide demonstrated co-localization with TG antigen. Transglutaminase activity was found in 6 of 14 coronary artery atherectomy samples. Cross-linking of TG substrates such as fibrinogen, fibronectin, vitronectin, collagen type I, and protease inhibitors stabilized the plaque. Furthermore, the activation of transforming growth factor-beta-1 by TG might be an additional mechanism for the promotion of plaque stabilization and progression by increasing the synthesis of extracellular matrix components. (Lab Invest 2001, 81:83-93).

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Transamidase kinetics. Amide formation in the enzymic reactions of thiol esters with amines

Cited by 45 publications

References 24 publications

Conserved tryptophan in the core domain of transglutaminase is essential for catalytic activity

Conserved tryptophan in the core domain of transglutaminase is essential for catalytic activity

Tissue Transglutaminase Mediates Activation of RhoA and MAP Kinase Pathways during Retinoic Acid-induced Neuronal Differentiation of SH-SY5Y Cells

Localization of Tissue Transglutaminase in Human Carotid and Coronary Artery Atherosclerosis: Implications for Plaque Stability and Progression

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