Biochemical characterization of a Ca2+/NAD(P)H-dependent H2O2 generator in human thyroid tissue

Leseney, Anne-Marie; Dème, Danielle; Legué, Odette; Ohayon, Renée; Chanson, Philippe; Sales, Jean-Patrick; Carvalho, Denise P.; Dupuy, Corinne; Virion, Alain

doi:10.1016/s0300-9084(99)80084-4

Cited by 60 publications

(64 citation statements)

References 29 publications

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“…Fig. 1A shows that the particulate fractions of human DUOX1-or DUOX2-transfected cells incubated with NADPH generated H 2 O 2 in a Ca 2ϩ -dependent manner, as did the thyroid particulate fraction (34). Interestingly, the activity of the particulate fraction from DUOX1 stably transfected cells was much lower than that found for their DUOX2 counterpart.…”

Section: Nadph/ca 2ϩ -Dependent H 2 O 2 Generationmentioning

confidence: 87%

“…They could trigger the reversible activation of the thyroid H 2 O 2 generator by calcium observed in vitro with porcine and human thyroid membrane fractions (34,38) and in intact cells from thyroid follicles (39) and slices (43,44). Recently, it has been clearly demonstrated that Nox5, which contains four calcium-binding sites in its N-terminal domain, including three canonical EF-hand motifs, is also directly activated by calcium (8).…”

Section: Nadph/ca 2ϩ -Dependent H 2 O 2 Generationmentioning

confidence: 99%

“…The NADPH-dependent H 2 O 2 -generating activity of the particles was determined as previously described (34). Samples of cellular particulate fraction were incubated at 30°C in 0.5 ml of 200 mM sodium phosphate, pH 7.4, containing 1 mM NaN 3 , 1 mM EGTA, 1.5 mM CaCl 2 , and 0.1 M FAD.…”

Section: Measurement Of the H 2 O 2 Production Of The Particulate Framentioning

confidence: 99%

“…Thyroid NADPH oxidase requires the presence of micromolar concentrations of calcium to acquire a functional conformation and to generate H 2 O 2 (34,37,38). Duox1 and Duox2 contain two Ca 2ϩ -binding motifs (EF-hand), which could be involved in the direct activation of the H 2 O 2 generator by calcium.…”

Section: Nadph/ca 2ϩ -Dependent H 2 O 2 Generationmentioning

confidence: 99%

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Dual Oxidase-2 Has an Intrinsic Ca2+-dependent H2O2-generating Activity

Ameziane-El-Hassani

Morand

Boucher

et al. 2005

Journal of Biological Chemistry

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208

155

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Duox2 (and probablyDuox1Reactive oxygen species (ROS) 1 have emerged as important molecules involved in regulating essential cell functions, such as growth and differentiation (1). NAD(P)H oxidases are a major source of ROS. Phagocyte oxidase is the oxidase that has been investigated most thoroughly (2). It catalyzes the production of superoxide by the one-electron reduction of oxygen, using NADPH as the electron donor. The catalytic moiety of the phagocyte NADPH oxidase is gp91phox , a plasma membraneassociated flavohemoprotein. Recently, it was discovered that gp91 phox belongs to a family consisting of several very similar oxidases. Seven NOX (NADPH oxidase) and DUOX/ThOX (dual oxidase/thyroid oxidase) genes have been identified that encode different NADPH oxidases with differing mRNA tissue expression. The Nox family comprises gp91phox , now known as Nox2; Nox1, which is predominantly expressed in the colon (3); Nox3, cloned from fetal kidney (4); Nox4 found in the kidney cortex (5, 6); and Nox5, expressed in the testis, spleen, and lymph nodes (7). In addition to the basic structure of gp91 phox , Nox5 has a long intracellular N-terminal domain with four calcium binding sites implicated in its Ca 2ϩ -dependent activation (8).The biological functions of these Nox proteins are now under investigation. They are involved in signal transduction related to cell growth and cancer (9 -11) and to angiogenesis (12).Duox1 and Duox2 are large homologues of Nox2 with an N-terminal extension comprising two EF-hand motifs, an additional transmembrane helix, and a peroxidase homology ectodomain (see Fig. 4A) (13,14). DUOX genes have been identified in the thyroid gland, where they are strongly expressed (13,14). However, the DUOX are also expressed on the mucosal surfaces of the trachea and the bronchi (15) and in the airway epithelial cells (16,17), where it has been suggested that Duox1 is the isoform responsible for acid production and secretion in airways (16) and plays a critical role in mucin expression (18). DUOX2 was also expressed throughout the digestive tract, where it was found to be functional (19,20), in addition to the salivary gland and rectum (15).It has been suggested that Duox2, which was identified by purifying thyroid NADPH oxidase, may constitute the catalytic core of this enzyme and generate the H 2 O 2 used by Tpo to catalyze the biosynthesis of thyroid hormones at the apical surface of the thyrocytes (13). Although no functional Duoxbased H 2 O 2 -generating system has yet been reconstituted (21), this proposal is corroborated by a recent report of permanent and severe congenital hypothyroidism in a patient with a bial-* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.¶ Recipient of a fellowship from the Commissariat à l'Energie Atomique (Paris, France).ʈ Recipients of a fellowship from the National Education, Research, an...

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Section: Nadph/ca 2ϩ -Dependent H 2 O 2 Generationmentioning

confidence: 87%

Section: Nadph/ca 2ϩ -Dependent H 2 O 2 Generationmentioning

confidence: 99%

Section: Measurement Of the H 2 O 2 Production Of The Particulate Framentioning

confidence: 99%

Section: Nadph/ca 2ϩ -Dependent H 2 O 2 Generationmentioning

confidence: 99%

See 2 more Smart Citations

Dual Oxidase-2 Has an Intrinsic Ca2+-dependent H2O2-generating Activity

Ameziane-El-Hassani

Morand

Boucher

et al. 2005

Journal of Biological Chemistry

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208

155

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“…The C-terminal region, comprising the last six TMDs of Duox, displays high sequence similarities with the other members of the NAD(P)H oxidase family (9), indicating that it probably catalyzes the transfer of electrons across the membrane from NADPH to molecular oxygen. The large intracellular loop located in the middle part of Duox, between the first two TMDs, contains two potential calcium EF-hand binding motifs and is probably involved in the control by calcium of the conformation and activity of NADPH oxidase (10,11). The ϳ600-residue-long extracellular N-terminal part of the protein is in part homologous to animal heme peroxidases and distinguishes the Duox family from other NAD(P)H oxidases.…”

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confidence: 99%

Targeting of the Dual Oxidase 2 N-terminal Region to the Plasma Membrane

Morand

Agnandji

Noël-Hudson

et al. 2004

Journal of Biological Chemistry

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-generating system has yet been obtained after transfecting Duox2 into non-thyroid cell lines, because it is retained in the endoplasmic reticulum (ER). We investigated the level of maturation of various Duox2 truncated proteins in an attempt to identify the region of Duox2 responsible for its remaining in the ER. Duox2-Q686X mutant, corresponding to the Nterminal ectodomain including the first putative transmembrane domain, was expressed in different cell lines. Carbohydrate content analysis revealed that complex type-specific Golgi apparatus (GA) oligosaccharides were present on pig Duox2-Q686X, whereas human truncated Duox2 carried only high mannose-type sugar chains characteristic of the ER. Further characterization using surface biotinylation and flow cytometry assays indicated that pig Duox2-Q686X was present at the plasma membrane, whereas human Duox2-Q686X remained inside the cell. The replacement of the last 90 residues of the human Duox2-Q686X with the pig equivalent region allowed the chimerical peptide to reach the Golgi apparatus. Pig mutants containing the complete first intracellular loop with or without the second transmembrane domain accumulated in the ER. These findings show that 1) the human Duox2-Q686X region encompassing residues 596 -685 prevents mutant exportation from the ER and 2) there is a pig Duox2 retention domain in the first intracellular loop. In addition, missense mutations of four cysteines (Cys-351, -370, -568, or -582) completely inhibited the emergence of pig Duox2-Q686X from the ER compartment, indicating their importance in Duox2 maturation.

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Effect of oxidative stress on rat thyrocyte iodide metabolism

Nadolnik

Niatsetskaya

Lupachyk

2007

Cell Biochemistry & Function

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Thyroid cells fall into the type of cells functioning during continuous production of high H(2)O(2) concentrations. We studied the effect of H(2)O(2)-induced oxidative stress (0.1, 1.0 and 10.0 mM) on the activities of the key steps of iodide metabolism (uptake, oxidation and organification) in thyrocytes cultivated in an organ culture. After 60 min cultivation of cells in a medium containing H(2)O(2) at concentrations of 1.0 and 10.0 mM iodide (I(-)) uptake, thyroperoxidase (TPO) activity and I(-) organification were completely inhibited. No restoration of the parameters studied was observed within the subsequent 24 h of cultivation. The inhibitory effect of 0.1 mM H(2)O(2) was reversible. Activation of I(-) uptake in the cultivated tissue and a 520-880% increase of the total I(-) content were observed after 8 and 24 h. The concentration of I(-) protein-bound fraction was raised by 220% after 24 h. A biphasic effect of 0.1 mM H(2)O(2) on TPO was observed: 76.2% and 72.2% inhibitions were seen after 2 and 8 h, respectively, whereas 40.0% enzyme activation was after 5 h. TPO activity was partially restored after 24 h and amounted to 65% of the initial value. The significant increase in the concentration of iodide protein-bound fraction, which was observed simultaneously with TPO inhibition, could be due to thyroglobulin non-enzymic iodination under H(2)O(2)-generated oxidative stress. The data obtained indicate that iodide oxidation, as a step in the biosynthesis of thyroid hormones, was most sensitive to oxidative stress activation. The impaired iodide uptake and its organification during oxidative stress can play a pathogenetic role in disturbed functions of thyroid cells.

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Biochemical characterization of a Ca2+/NAD(P)H-dependent H2O2 generator in human thyroid tissue

Cited by 60 publications

References 29 publications

Dual Oxidase-2 Has an Intrinsic Ca2+-dependent H2O2-generating Activity

Dual Oxidase-2 Has an Intrinsic Ca2+-dependent H2O2-generating Activity

Targeting of the Dual Oxidase 2 N-terminal Region to the Plasma Membrane

Effect of oxidative stress on rat thyrocyte iodide metabolism

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