Molecular Cloning and Functional Expression of the Gene Encoding the Human Proteinase‐Activated Receptor 2
- 1 August 1995
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 232 (1) , 84-89
- https://doi.org/10.1111/j.1432-1033.1995.tb20784.x
Abstract
We previously reported the molecular cloning of a mouse guanosine-nucleotide-binding-protein-coupled receptor similar to the thrombin receptor. Since the physiological agonist was unknown, the receptor was named proteinase-activated receptor 2. We describe here the cloning and functional expression of the gene encoding the corresponding human receptor. The gene is divided into two exons separated by about 14 kb intronic DNA. The deduced protein sequence is 397 amino acids long and 83% identical to the mouse receptor sequence. Within the extracellular amino terminus, the residues predicted to form the tethered agonist ligand differ between the two receptors; of the first six residues only four are conserved. At positions five and six, a lysine residue and a valine residue, respectively, have replaced arginine and leucine residues found in the mouse sequence. When the human receptor is expressed in Chinese hamster ovary cells, it can be activated by low nanomolar concentrations of the serine proteinase trypsin and by peptides made from the receptor sequence. Northern-blot analysis of receptor expression showed that the receptor transcript is widely expressed in human tissues with especially high levels in pancreas, liver, kidney, small intestine and colon. Moderate expression was detected in many organs but none in brain or skeletal muscle. By fluorescence in situ hybridization, the human proteinase-activated receptor 2 gene was mapped to chromosomal region 5q13, where, previously, the related thrombin receptor gene has been located.Keywords
This publication has 18 references indexed in Scilit:
- The Mouse Proteinase-activated Receptor-2 cDNA and GeneJournal of Biological Chemistry, 1995
- α-thrombin and trypsin use different receptors to stimulate arachidonic acid metabolismProstaglandins, 1994
- Specificity of the thrombin receptor for agonist peptide is defined by its extracellular surfaceNature, 1994
- Essential groups in synthetic agonist peptides for activation of the platelet thrombin receptorBiochemistry, 1992
- Sequence Alignment of the G-Protein Coupled Receptor SuperfamilyDNA and Cell Biology, 1992
- Domains specifying thrombin–receptor interactionNature, 1991
- cDNA cloning and expression of a hamster α‐thrombin receptor coupled to Ca2+ mobilizationFEBS Letters, 1991
- Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activationCell, 1991
- Tandem Array of Human Visual Pigment Genes at Xq28Science, 1988
- Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomesCell, 1986