Cloning and characterization of the ecto-nucleotidase NTPDase3 from rat brain: Predicted secondary structure and relation to other members of the E-NTPDase family and actin.

Purinergic Signalling
Thomas VorhoffNorbert Braun

Abstract

The protein family of ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDase family) contains multiple members that hydrolyze nucleoside 5'-triphosphates and nucleoside 5'-diphosphates with varying preference for the individual type of nucleotide. We report the cloning and functional expression of rat NTPDase3. The rat brain-derived cDNA has an open reading frame of 1590 bp encoding 529 amino acid residues, a calculated molecular mass of 59.1 kDa and predicted N- and C-terminal hydrophobic sequences. It shares 94.3% and 81.7% amino acid identity with the mouse and human NTPDase3, respectively, and is more closely related to cell surface-located than to the intracellularly located members of the enzyme family. The NTPDase3 gene is allocated to chromosome 8q32 and organized into 11 exons. Rat NTPDase3 expressed in CHO cells hydrolyzed both nucleoside triphosphates and nucleoside diphosphates with hydrolysis ratios of ATP:ADP of 5:1 and UTP:UDP of 8:1. After addition of ATP, ADP is formed as an intermediate product that is further hydrolyzed to AMP. The enzyme is preferentially activated by Ca(2+) over Mg(2+) and reveals an alkaline pH optimum. Immunocytochemistry confirmed expression of heterologously expressed NTPDase3 to t...Continue Reading

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Citations

Apr 13, 2011·Purinergic Signalling·Aileen F Knowles
May 5, 2012·Purinergic Signalling·Herbert ZimmermannNorbert Sträter
Mar 17, 2007·Life Sciences·Fernanda Cenci VuadenCarla Denise Bonan
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Mar 9, 2013·Plant Signaling & Behavior·Riku OkuhataNobuyuki Kanzawa
May 7, 2008·Proceedings of the National Academy of Sciences of the United States of America·Matthias Zebisch, Norbert Sträter
Jan 1, 2009·Synapse·María Jimena RicattiMaría Paula Faillace
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Mar 2, 2006·Pharmacological Reviews·Geoffrey Burnstock
Jan 15, 2011·American Journal of Physiology. Gastrointestinal and Liver Physiology·Elise G LavoieJean Sévigny
Jun 26, 2013·Molecular and Cellular Biochemistry·Fátima Husein AbdallaCinthia Melazzo Mazzanti
Aug 2, 2017·Chemical Communications : Chem Comm·Atara Nardi-SchreiberRachel Katz-Brull
Aug 5, 2010·American Journal of Physiology. Endocrinology and Metabolism·Elise G LavoieJean Sévigny
Nov 21, 2020·Psychiatry Research·Carolina GubertAna Maria Oliveira Battastini
Apr 22, 2021·The Journal of Immunology : Official Journal of the American Association of Immunologists·Clarissa Branco HaasGyörgy Haskó
Oct 7, 2018·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Naiara StefanelloMaria Rosa Chitolina Schetinger
Oct 1, 2021·Purinergic Signalling·Andréia Machado CardosoSarah Franco Vieira de Oliveira Maciel

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Datasets Mentioned

BETA
AF034840
AJ437217

Methods Mentioned

BETA
PCR
electrophoresis
glycosylation
transfection
phosphotransferase

Software Mentioned

TMHMM
BoxShade
Omiga
SignalP
BLAST
TreeView
ClustalX
SSpro

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