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Molecular cloning of the rat proteinase-activated receptor 4 (PAR4).

Hoogerwerf WA, Hellmich HL, Micci MA, Winston JH, Zou L, Pasricha PJ - BMC Mol. Biol. (2002)

Bottom Line: We 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Enteric Neuromuscular Disorders and Pain Laboratory, Division of Gastroenterology and Hepatology, University of Texas Medical Branch, Galveston, TX, USA. wahooger@utmb.edu

ABSTRACT

Background: The 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.

Results: The 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 microM) and a putative rat PAR4 specific activating peptide (100 microM), as measured by intracellular Ca2+-changes.

Conclusions: We 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.

No MeSH data available.


Northern blot of rat duodenal mRNA hybridized to a rat PAR4 probe. Left: ribosomal markers. The single hybridizing band corresponds to a mRNA of ≅ 3 kb.
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Figure 3: Northern blot of rat duodenal mRNA hybridized to a rat PAR4 probe. Left: ribosomal markers. The single hybridizing band corresponds to a mRNA of ≅ 3 kb.

Mentions: Northern blot analysis of poly(A)+ RNA from the duodenum detected a 3.0 kb band (Figure 3).


Molecular cloning of the rat proteinase-activated receptor 4 (PAR4).

Hoogerwerf WA, Hellmich HL, Micci MA, Winston JH, Zou L, Pasricha PJ - BMC Mol. Biol. (2002)

Northern blot of rat duodenal mRNA hybridized to a rat PAR4 probe. Left: ribosomal markers. The single hybridizing band corresponds to a mRNA of ≅ 3 kb.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC88883&req=5

Figure 3: Northern blot of rat duodenal mRNA hybridized to a rat PAR4 probe. Left: ribosomal markers. The single hybridizing band corresponds to a mRNA of ≅ 3 kb.
Mentions: Northern blot analysis of poly(A)+ RNA from the duodenum detected a 3.0 kb band (Figure 3).

Bottom Line: We 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Enteric Neuromuscular Disorders and Pain Laboratory, Division of Gastroenterology and Hepatology, University of Texas Medical Branch, Galveston, TX, USA. wahooger@utmb.edu

ABSTRACT

Background: The 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.

Results: The 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 microM) and a putative rat PAR4 specific activating peptide (100 microM), as measured by intracellular Ca2+-changes.

Conclusions: We 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.

No MeSH data available.