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Your search keyword '"Pouillon, V."' showing total 2 results
Start Over Author "Pouillon, V." Topic cloning, molecular
2 results on '"Pouillon, V."'

1. Molecular cloning and characterization of the mouse P2Y4 nucleotide receptor.

Author

Suarez-Huerta N, Pouillon V, Boeynaems J, and Robaye B

Subjects
Adenosine Triphosphate pharmacology, Amino Acid Sequence, Animals, Antineoplastic Agents pharmacology, Dose-Response Relationship, Drug, Humans, Inositol 1,4,5-Trisphosphate metabolism, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Platelet Aggregation Inhibitors pharmacology, Pyridoxal Phosphate analogs & derivatives, Pyridoxal Phosphate pharmacology, Rats, Receptors, Purinergic P2 chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Sequence Alignment, Suramin pharmacology, Transfection, Triazines pharmacology, Tumor Cells, Cultured, Uridine Triphosphate pharmacology, Cloning, Molecular, Receptors, Purinergic P2 genetics, Receptors, Purinergic P2 metabolism
Abstract

To isolate the mouse P2Y4 receptor gene, a mouse genomic library was screened with a human P2Y4 probe. An open reading frame encoding a protein of 361 amino acids was isolated. This protein showed 82% and 95% amino acid identity with the human and rat P2Y4 receptors, respectively. By reverse transcription and polymerase chain reaction (RT-PCR), the P2Y4 messenger RNA was detected in mouse liver, intestine, stomach, bladder and lung among the 16 mouse tissues tested. In 1321N1 transfected cells, the mouse P2Y4 receptor was equally activated by UTP and ATP, and was antagonized by pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) and Reactive Blue 2, and not by suramin. Moreover, when expressed in 1321N1 cells, the rat P2Y4 is also antagonized by PPADS. Thus, when compared in the same expression system, the mouse P2Y4 is closer to the rat ortholog in terms of agonist stimulation, while in terms of antagonist profile, the three P2Y4 receptor orthologs are similar.

Published
2001
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2. Cloning and expression of a cDNA encoding human inositol 1,4,5-trisphosphate 3-kinase C

Author

Dewaste, V, Pouillon, V, Moreau, C, Shears, S, Takazawa, K, and Erneux, C

Subjects
DNA, Complementary, Base Sequence, Sequence Homology, Amino Acid, Molecular Sequence Data, Recombinant Proteins, Phosphotransferases (Alcohol Group Acceptor), COS Cells, Animals, Humans, Calcium, Amino Acid Sequence, Cloning, Molecular, Chromatography, High Pressure Liquid, Research Article, DNA Primers
Abstract

Inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] 3-kinase catalyses the phosphorylation of Ins(1,4,5)P(3) to Ins(1,3,4,5)P(4). cDNAs encoding two isoenzymes of Ins(1,4,5)P(3) 3-kinase (3-kinases A and B) have been described previously. In the present study, we report the cloning of a full-length 2052 bp cDNA encoding a third human isoenzyme of the Ins(1,4,5)P(3) 3-kinase family, referred to as isoform C. This novel enzyme has a calculated molecular mass of 75. 207 kDa and a K(m) for Ins(1,4,5)P(3) of 6 microM. Northern-blot analysis showed the presence of a transcript of approx. 3.9 kb in various human tissues. Inositol trisphosphate 3-kinase C demonstrates enzymic activity when expressed in DH5alphaF' bacteria or COS-7 cells. Calcium alone decreases the Ins(1,4,5)P(3) 3-kinase activity of the 3-kinase C isoenzyme in transfected COS-7 cells. This inhibitory effect is reversed in the presence of calmodulin. The recombinant bacterial 3-kinase C can be adsorbed on calmodulin-Sepharose in the presence of calcium. The present data show that Ins(1,4,5)P(3) 3-kinase C: (i) shares a conserved catalytic domain of about 275 amino acids with the two other mammalian isoforms, (ii) could be purified on a calmodulin-Sepharose column and (iii) could be distinguished from the A and B isoenzymes by the effects of calcium and of calmodulin.

Published
2000

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