Colocalization of Tissue Transglutaminase and Stress Fibers in Human Vascular Smooth Muscle Cells and Human Umbilical Vein Endothelial Cells

Chowdhury, Zinnat A.; Barsigian, Carl; Chalupowicz, Graciela D.; Bach, Tami L.; Garcia‐Manero, Guillermo; Martinez, Josè

doi:10.1006/excr.1996.3448

Cited by 45 publications

(32 citation statements)

References 42 publications

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“…This strongly suggests that plasma membrane-associated Gh␣ is accessible to the interaction with G protein-coupled receptors in pregnant rat myometrium. Previous findings in human vascular smooth muscle cells reported that the particulate Gh␣ codistributes with stress fibers and may thus stabilize cytoskeletal structures through its cross-linking function (42). In the latter study, the particulate Gh␣ appeared inactive when assayed by in vitro putrescine/casein assay, maybe due to its tight binding to preferred local substrates (42).…”

Section: Expression Of Gh␣ In Rat Myometrium-by Using Rt-pcrmentioning

confidence: 74%

“…Previous findings in human vascular smooth muscle cells reported that the particulate Gh␣ codistributes with stress fibers and may thus stabilize cytoskeletal structures through its cross-linking function (42). In the latter study, the particulate Gh␣ appeared inactive when assayed by in vitro putrescine/casein assay, maybe due to its tight binding to preferred local substrates (42). It is known that stimulation of transglutaminase activity into cells requires both high concentrations of Ca 2ϩ and a decrease of guanine nucleotide levels (43,44).…”

Section: Expression Of Gh␣ In Rat Myometrium-by Using Rt-pcrmentioning

confidence: 83%

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Functional Coupling of Rat Myometrial α1-Adrenergic Receptors to Ghα/Tissue Transglutaminase 2 during Pregnancy

Morgan

Levy

Mhaouty-Kodja

2004

Journal of Biological Chemistry

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Gh␣ protein, which exhibits both transglutaminase and GTPase activities, represents a new class of GTPbinding proteins. In the present study, we characterized Gh␣ in rat uterine smooth muscle (myometrium) and followed its expression during pregnancy by reverse transcription-PCR and Western blot. We also measured transglutaminase and GTP binding functions and used a smooth muscle cell line to evaluate the role of Gh␣ in cell proliferation. The results show that pregnancy is associated with an up-regulation of Gh␣ expression at both the mRNA and protein level. Gh␣ induced during pregnancy is preferentially localized to the plasma membrane. This was found associated with an increased ability of plasma membrane preparations to catalyze Ca 2؉ -dependent incorporation of [ 3 H]putrescine into casein in vitro. In the cytosol, significant changes in the level of immunodetected Gh␣ and transglutaminase activity were seen only at term. Activation of ␣1-adrenergic receptors (␣1-AR) enhanced photoaffinity labeling of plasma membrane Gh␣. Moreover, the level of ␣1-ARcoupled Gh␣ increased progressively with pregnancy, which parallels the active period of myometrial cell proliferation. Overexpression of wild type Gh␣ in smooth muscle cell line DDT1-MF2 increased ␣1-AR-induced [ 3 H]thymidine incorporation. A similar response was obtained in cells expressing the transglutaminase inactive mutant (C277S) of Gh␣. Together, these findings underscore the role of Gh␣ as signal transducer of ␣1-ARinduced smooth muscle cell proliferation. In this context, pregnant rat myometrium provides an interesting physiological model to study the mechanisms underlying the regulation of the GTPase function of Gh␣.

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Section: Expression Of Gh␣ In Rat Myometrium-by Using Rt-pcrmentioning

confidence: 74%

Section: Expression Of Gh␣ In Rat Myometrium-by Using Rt-pcrmentioning

confidence: 83%

Functional Coupling of Rat Myometrial α1-Adrenergic Receptors to Ghα/Tissue Transglutaminase 2 during Pregnancy

Morgan

Levy

Mhaouty-Kodja

2004

Journal of Biological Chemistry

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“…Human arterial SMCs showed a 3-fold increase in tTG activity with atRA stimulation, but the mRNA expression of tTG was not reported in these cells. 29 Interestingly, tTG colocalized with stress fibers in human arterial and venous SMCs and was shown to coimmunoprecipitate with myosin, suggesting that codistribution of tTG with stress fibers was attributable to tTG's interaction with myosin. 29 Although we did observe some evidence of stress fiber formation in atRA-treated SMCs, which is consistent with atRA's prodifferentiation effects in these cells, extensive confocal microscopy failed to reveal any colocalization of tTG with the stress fiber apparatus (data not shown).…”

Section: Discussionmentioning

confidence: 92%

Retinoic Acid–Induced Tissue Transglutaminase and Apoptosis in Vascular Smooth Muscle Cells

Haendeler

Aebly

et al. 2000

Circulation Research

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Abstract-Retinoids exert antiproliferative and prodifferentiating effects in vascular smooth muscle cells (SMCs) and reduce neointimal mass in balloon-injured blood vessels. The mechanisms through which retinoids carry out these effects are unknown but likely involve retinoid receptor-mediated changes in gene expression. Here we report the cloning, chromosomal mapping, and biological activity of the retinoid-response gene rat tissue transglutaminase (tTG).Northern blotting studies showed that tTG is rapidly and dose-dependently induced in a protein synthesis-independent manner after stimulation with the natural retinoid all-trans retinoic acid (atRA). The induction of tTG was selective for atRA and its stereoisomers 9-cis and 13-cis RA, because little or no elevation in mRNA expression was observed with a panel of growth factors. Western blotting and immunofluorescence confocal microscopy showed an accumulation of cytosolic tTG protein after atRA stimulation. Radiolabeled cross-linking studies revealed a corresponding elevation in in vitro tTG activity. The increase in tTG activity was reduced in the presence of 2 distinct inhibitors of tTG (monodansylcadaverine and cystamine). atRA-induced tTG mRNA and protein expression were followed by a significant elevation in SMC apoptosis. Such retinoid-induced programmed cell death could be partially inhibited with each tTG inhibitor and was completely blocked when both inhibitors were used simultaneously. These results establish a role for atRA in the sequential stimulation of tTG and apoptosis in cultured SMCs. atRA-mediated apoptosis in SMCs seems to require the participation of active tTG, suggesting a potential mechanistic link between this retinoid-inducible gene and programmed cell death. (Circ Res. 2000;87:881-887.)

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“…Moreover, it has been reported that tTG is associated with elements of the cytoskeleton, thus indicating its involvement in the organization of the cytoskeleton. tTG was in fact shown to be co-localized with the stress fibers via cross-linking to myosin by virtue of autocatalytic activity of the enzyme (41). In this study, we report that ezrin/radixin/moesin-binding phosphoprotein-50, a member of ezrin/radixin/moesin family of membrane-cytoskeletal adapters involved in the formation of microvilli, cell-cell adhesion, maintenance of cell shape, cell motility, and membrane trafficking (42), is able to act as a tTG Lys-donor substrate.…”

Section: Discussionmentioning

confidence: 99%

Proteomics Identification of Acyl-acceptor and Acyl-donor Substrates for Transglutaminase in a Human Intestinal Epithelial Cell Line

Orrú

Caputo

D’Amato

et al. 2003

Journal of Biological Chemistry

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Transglutaminase (TG)-catalyzed cross-linking of both intracellular and extracellular proteins is an important biochemical event. However, increased concentrations of cross-linked proteins have been observed in many disorders. Moreover, TG-catalyzed modification of proteins might generate new self-antigens responsible for the autoimmune response, as in celiac disease. The identification of available substrates may offer an understanding of how the TG-catalyzed post-translational modification has an impact on physiology and disease. We used a proteomic approach to identify TG-modified protein targets in human intestinal epithelial cells to determine the extent to which transglutaminase specifically contributes to celiac disease. Two probes were used for endogenous TG activity: 5-(biotinamido)pentylamine, which represents the acyl-acceptor, and a biotinylated glutamine-containing peptide, which represents the acyl-donor. This approach identified >25 proteins, which range from 30,000 to 300,000 Daltons and can serve as acyl-acceptor and/or acyl-donor for transglutaminase. Some of them were known transglutaminase substrates, whereas others had not been previously identified. These targets include proteins involved in cytoskeletal network organization, folding of proteins, transport processes, and miscellaneous metabolic functions.

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Colocalization of Tissue Transglutaminase and Stress Fibers in Human Vascular Smooth Muscle Cells and Human Umbilical Vein Endothelial Cells

Cited by 45 publications

References 42 publications

Functional Coupling of Rat Myometrial α1-Adrenergic Receptors to Ghα/Tissue Transglutaminase 2 during Pregnancy

Functional Coupling of Rat Myometrial α1-Adrenergic Receptors to Ghα/Tissue Transglutaminase 2 during Pregnancy

Retinoic Acid–Induced Tissue Transglutaminase and Apoptosis in Vascular Smooth Muscle Cells

Proteomics Identification of Acyl-acceptor and Acyl-donor Substrates for Transglutaminase in a Human Intestinal Epithelial Cell Line

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