Cloning, molecular characterization and expression of ecto-nucleoside triphosphate diphosphohydrolase-1 from Torpedo electric organ

2007 ◽  
Vol 50 (1) ◽  
pp. 256-263 ◽  
Author(s):  
Mireia Martín-Satué ◽  
Benjamín Torrejón-Escribano ◽  
Antonio Felipe ◽  
Inmaculada Gómez de Aranda ◽  
Marc Elías ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Electric Organ ◽  
Nucleoside Triphosphate ◽  
Nucleoside Triphosphate Diphosphohydrolase ◽  
Torpedo Electric Organ

Gadolinium Inhibition of Ecto-Nucleoside Triphosphate Diphosphohydrolase Activity in Torpedo Electric Organ

2004 ◽  
Vol 29 (9) ◽  
pp. 1711-1714 ◽  
Author(s):  
Artur Escalada ◽  
Piedad Navarro ◽  
Esteve Ros ◽  
Jordi Aleu ◽  
Carles Solsona ◽  
...  
Keyword(s):  
Electric Organ ◽  
Nucleoside Triphosphate ◽  
Nucleoside Triphosphate Diphosphohydrolase ◽  
Torpedo Electric Organ

scholarly journals NTPDase1 Modulates Smooth Muscle Contraction in Mice Bladder by Regulating Nucleotide Receptor Activation Distinctly in Male and Female

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 147
Author(s):  
Romuald Brice Babou Kammoe ◽  
Gilles Kauffenstein ◽  
Julie Pelletier ◽  
Bernard Robaye ◽  
Jean Sévigny
Keyword(s):  
Smooth Muscle ◽  
Ex Vivo ◽  
Smooth Muscle Contraction ◽  
Receptor Activation ◽  
Nucleoside Triphosphate ◽  
Nucleotide Receptors ◽  
Nerve Terminals ◽  
Wild Type ◽  
Nucleoside Triphosphate Diphosphohydrolase ◽  
Bladder Contraction

Nucleotides released by smooth muscle cells (SMCs) and by innervating nerve terminals activate specific P2 receptors and modulate bladder contraction. We hypothesized that cell surface enzymes regulate SMC contraction in mice bladder by controlling the concentration of nucleotides. We showed by immunohistochemistry, enzymatic histochemistry, and biochemical activities that nucleoside triphosphate diphosphohydrolase-1 (NTPDase1) and ecto-5′-nucleotidase were the major ectonucleotidases expressed by SMCs in the bladder. RT-qPCR revealed that, among the nucleotide receptors, there was higher expression of P2X1, P2Y1, and P2Y6 receptors. Ex vivo, nucleotides induced a more potent contraction of bladder strips isolated from NTPDase1 deficient (Entpd1−/−) mice compared to wild type controls. The strongest responses were obtained with uridine 5′-triphosphate (UTP) and uridine 5′-diphosphate (UDP), suggesting the involvement of P2Y6 receptors, which was confirmed with P2ry6−/− bladder strips. Interestingly, this response was reduced in female bladders. Our results also suggest the participation of P2X1, P2Y2 and/or P2Y4, and P2Y12 in these contractions. A reduced response to the thromboxane analogue U46619 was also observed in wild type, Entpd1−/−, and P2ry6−/− female bladders showing another difference due to sex. In summary, NTPDase1 modulates the activation of nucleotide receptors in mouse bladder SMCs, and contractions induced by P2Y6 receptor activation were weaker in female bladders.

scholarly journals Diadenosine polyphosphate hydrolase from presynaptic plasma membranes of Torpedo electric organ

1997 ◽  
Vol 323 (3) ◽  
pp. 677-684 ◽  
Author(s):  
Jesús MATEO ◽  
Pedro ROTLLAN ◽  
Eulalia MARTI ◽  
Inmaculada GOMEZ DE ARANDA ◽  
Carles SOLSONA ◽  
...  
Keyword(s):  
Plasma Membranes ◽  
Pyridoxal Phosphate ◽  
Electric Organ ◽  
Competitive Inhibitor ◽  
Inhibitory Effect ◽  
Ic50 Value ◽  
Neural Origin ◽  
Disulphonic Acid ◽  
Torpedo Electric Organ ◽  
Enzyme Activators

The diadenosine polyphosphate hydrolase present in presynaptic plasma membranes from the Torpedo electric organ has been characterized using fluorogenic substrates of the form di-(1,N6-ethenoadenosine) 5´,5‴-P1,Pn-polyphosphate. The enzyme hydrolyses diadenosine polyphosphates (Apn A, where n = 3–5), producing AMP and the corresponding adenosine (n-1) 5´-phosphate, Ap(n-1). The Km values of the enzyme were 0.543± 0.015, 0.478±0.043 and 0.520±0.026 μM, and the Vmax values were 633±4, 592±18 and 576±45 pmol/min per mg of protein, for the etheno derivatives of Ap3A (adenosine 5´,5‴-P1,P3-triphosphate), Ap4A (adenosine 5´,5‴-P1,P4 -tetraphosphate) and Ap5A (adenosine 5´,5‴-P1,P5-pentaphosphate) respectively. Ca2+, Mg2+ and Mn2+ are enzyme activators, with EC50 values of 0.86±0.11, 1.35±0.24 and 0.58±0.10 mM respectively. The fluoride ion is an inhibitor with an IC50 value of 1.38±0.19 mM. The ATP analogues adenosine 5´-tetraphosphate and adenosine 5´-[γ-thio]triphosphate are potent competitive inhibitors and adenosine 5´-[α,β-methylene]diphosphate is a less potent competitive inhibitor, the Ki values being 0.29±0.03, 0.43±0.05 and 7.18±0.8 μM respectively. The P2-receptor antagonist pyridoxal phosphate 6-azophenyl-2´,4´-disulphonic acid behaves as a non-competitive inhibitor with a Ki value of 29.7±3.1 μM, and also exhibits a significant inhibitory effect on Torpedo apyrase activity. The effect of pH on the Km and Vmax values, together with inhibition by diethyl pyrocarbonate, strongly suggests the presence of functional histidine residues in Torpedo diadenosine polyphosphate hydrolase. The enzyme from Torpedo shows similarities with that of neural origin from neurochromaffin cells, and significant differences compared with that from endothelial vascular cells.

scholarly journals Large-scale purification of presynaptic plasma membranes from Torpedo marmorata electric organ.

1985 ◽  
Vol 101 (5) ◽  
pp. 1757-1762 ◽  
Author(s):  
N Morel ◽  
J Marsal ◽  
R Manaranche ◽  
S Lazereg ◽  
J C Mazie ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Large Scale ◽  
Plasma Membranes ◽  
Electric Organ ◽  
Acetylcholinesterase Activity ◽  
Cholinergic Nerve Terminals ◽  
Membrane Bound ◽  
Torpedo Electric Organ ◽  
Presynaptic Plasma Membrane ◽  
Glycera Convoluta

The presynaptic plasma membrane (PSPM) of cholinergic nerve terminals was purified from Torpedo electric organ using a large-scale procedure. Up to 500 g of frozen electric organ were fractioned in a single run, leading to the isolation of greater than 100 mg of PSPM proteins. The purity of the fraction is similar to that of the synaptosomal plasma membrane obtained after subfractionation of Torpedo synaptosomes as judged by its membrane-bound acetylcholinesterase activity, the number of Glycera convoluta neurotoxin binding sites, and the binding of two monoclonal antibodies directed against PSPM. The specificity of these antibodies for the PSPM is demonstrated by immunofluorescence microscopy.

scholarly journals Role of Ca2+ in Mediating Plant Responses to Extracellular ATP and ADP

2018 ◽  
Vol 19 (11) ◽  
pp. 3590 ◽  
Author(s):  
Greg Clark ◽  
Stanley Roux
Keyword(s):  
Cytosolic Calcium ◽  
Defense Responses ◽  
Receptor Activation ◽  
Extracellular Atp ◽  
Extracellular Nucleotides ◽  
Nucleoside Triphosphate ◽  
Plant Responses ◽  
Downstream Signaling ◽  
Nucleoside Triphosphate Diphosphohydrolase

Among the most recently discovered chemical regulators of plant growth and development are extracellular nucleotides, especially extracellular ATP (eATP) and extracellular ADP (eADP). Plant cells release ATP into their extracellular matrix under a variety of different circumstances, and this eATP can then function as an agonist that binds to a specific receptor and induces signaling changes, the earliest of which is an increase in the concentration of cytosolic calcium ([Ca2+]cyt). This initial change is then amplified into downstream-signaling changes that include increased levels of reactive oxygen species and nitric oxide, which ultimately lead to major changes in the growth rate, defense responses, and leaf stomatal apertures of plants. This review presents and discusses the evidence that links receptor activation to increased [Ca2+]cyt and, ultimately, to growth and diverse adaptive changes in plant development. It also discusses the evidence that increased [Ca2+]cyt also enhances the activity of apyrase (nucleoside triphosphate diphosphohydrolase) enzymes that function in multiple subcellular locales to hydrolyze ATP and ADP, and thus limit or terminate the effects of these potent regulators.

scholarly journals Evolution and comparative genomics of subcellular specializations: EST sequencing of Torpedo electric organ

2011 ◽  
Vol 4 (1) ◽  
pp. 33-40 ◽  
Author(s):  
Javad Nazarian ◽  
Deborah L. Berry ◽  
Salar Sanjari ◽  
Mohammed Razvi ◽  
Kristy Brown ◽  
...  
Keyword(s):  
Comparative Genomics ◽  
Electric Organ ◽  
Est Sequencing ◽  
Torpedo Electric Organ
Sign in / Sign up

Export Citation Format

Share Document