Molecular cloning of the rat proteinase-activated receptor 4 PAR4Report as inadecuate

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BMC Molecular Biology

, 3:2

First Online: 18 February 2002Received: 26 November 2001Accepted: 18 February 2002


BackgroundThe proteinase-activated receptor 4 PAR4 is a G-protein-coupled receptor activated by proteases such as thrombin and trypsin. Although activation of PAR4 has been shown to modulate rat gastrointestinal motility, the rat PAR4 sequence was unknown until now. This study aimed to identify the rat PAR4 cDNA.

ResultsThe cDNA coding for the rat PAR4 homologue was cloned from the duodenum. Northern blots demonstrated a 3.0 kb transcript in the duodenum. Protein homology with mouse and human counterparts was 90% and 75% respectively. PAR4 is expressed predominantly in the esophagus, stomach, duodenum and the spleen. When expressed in COS cells, PAR4 is activated by trypsin 1 nM, thrombin 50 nM, mouse PAR4 specific peptide 500 μM and a putative rat PAR4 specific activating peptide 100 μM, as measured by intracellular Ca-changes.

ConclusionsWe have identified and characterized cDNA encoding the rat PAR4 homologue. PAR4 is expressed predominantly in the upper gastrointestinal tract. It is activated by trypsin, thrombin and its newly identified rat PAR4 specific activating peptide.

AbbreviationsPARproteinase-activated receptor

RPAribonuclease protection assay

APactivating peptide

ORFopen reading frame

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2199-3-2 contains supplementary material, which is available to authorized users.

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Author: Willemijntje A Hoogerwerf - Helen Lee Hellmich - Maria Adelaide Micci - John H Winston - Lei Zou - Pankaj J Pasricha


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