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A polymorphic protease activated receptor 2 (PAR2) displaying reduced sensitivity to trypsin and differential responses to PAR agonists

A polymorphic protease activated receptor 2 (PAR2) displaying reduced sensitivity to trypsin and differential responses to PAR agonists
A polymorphic protease activated receptor 2 (PAR2) displaying reduced sensitivity to trypsin and differential responses to PAR agonists
Protease-activated receptor 2 (PAR2) is a trypsin-activated member of a family of G-protein-coupled PARs. We have identified a polymorphic form of human PAR2 (PAR2F240S) characterized by a phenylalanine to serine mutation at residue 240 within extracellular loop 2, with allelic frequencies of 0.916 (Phe240) and 0.084 (Ser240) for the wild-type and mutant alleles, respectively.

Elevations in intracellular calcium were measured in permanently transfected cell lines expressing the receptors. PAR2F240S displayed a significant reduction in sensitivity toward trypsin (?3.7-fold) and the PAR2-activating peptides, SLIGKV-NH2 (?2.5-fold) and SLIGRL-NH2(?2.8-fold), but an increased sensitivity toward the selective PAR2 agonist,trans-cinnamoyl-LIGRLO-NH2(?4-fold). Increased sensitivity was also observed toward the selective PAR-1 agonist, TFLLR-NH2 (?7-fold), but not to other PAR-1 agonists tested. Furthermore, we found that TLIGRL-NH2 and a PAR4-derived peptide,trans-cinnamoyl-YPGKF-NH2, were selective PAR2F240S agonists.

By introducing the F240S mutation into rat PAR2, we observed shifts in agonist potencies that mirrored the human PAR2F240S, suggesting that Phe240 is involved in determining agonist specificity of PAR2. Finally, differences in receptor signaling were paralleled in a cell growth assay. We suggest that the distinct pharmacological profile induced by this polymorphism will have important implications for the design of PAR-targeted agonists/antagonists and may contribute to, or be predictive of, an inflammatory disease.
0021-9258
39207-39212
Compton, Steven J.
d86f4b0c-c58e-41be-9feb-1d1b0caebb04
Cairns, Jennifer A.
b74d6f9e-b4f0-423a-9dba-7a415a11f338
Palmer, Karan-Jane
931b90dc-6b20-4770-895b-55fe30d7346c
AL-Ani, Bahjat
b7079a92-b95b-416b-b8c3-710517d5c04d
Hollenberg, Morley D.
a186d69d-ba25-4dd6-81e5-6a94c04e7b35
Walls, Andrew F.
aaa7e455-0562-4b4c-94f5-ec29c74b1bfe
Compton, Steven J.
d86f4b0c-c58e-41be-9feb-1d1b0caebb04
Cairns, Jennifer A.
b74d6f9e-b4f0-423a-9dba-7a415a11f338
Palmer, Karan-Jane
931b90dc-6b20-4770-895b-55fe30d7346c
AL-Ani, Bahjat
b7079a92-b95b-416b-b8c3-710517d5c04d
Hollenberg, Morley D.
a186d69d-ba25-4dd6-81e5-6a94c04e7b35
Walls, Andrew F.
aaa7e455-0562-4b4c-94f5-ec29c74b1bfe

Compton, Steven J., Cairns, Jennifer A., Palmer, Karan-Jane, AL-Ani, Bahjat, Hollenberg, Morley D. and Walls, Andrew F. (2000) A polymorphic protease activated receptor 2 (PAR2) displaying reduced sensitivity to trypsin and differential responses to PAR agonists. The Journal of Biological Chemistry, 275, 39207-39212. (doi:10.1074/jbc.M007215200). (PMID:10995771)

Record type: Article

Abstract

Protease-activated receptor 2 (PAR2) is a trypsin-activated member of a family of G-protein-coupled PARs. We have identified a polymorphic form of human PAR2 (PAR2F240S) characterized by a phenylalanine to serine mutation at residue 240 within extracellular loop 2, with allelic frequencies of 0.916 (Phe240) and 0.084 (Ser240) for the wild-type and mutant alleles, respectively.

Elevations in intracellular calcium were measured in permanently transfected cell lines expressing the receptors. PAR2F240S displayed a significant reduction in sensitivity toward trypsin (?3.7-fold) and the PAR2-activating peptides, SLIGKV-NH2 (?2.5-fold) and SLIGRL-NH2(?2.8-fold), but an increased sensitivity toward the selective PAR2 agonist,trans-cinnamoyl-LIGRLO-NH2(?4-fold). Increased sensitivity was also observed toward the selective PAR-1 agonist, TFLLR-NH2 (?7-fold), but not to other PAR-1 agonists tested. Furthermore, we found that TLIGRL-NH2 and a PAR4-derived peptide,trans-cinnamoyl-YPGKF-NH2, were selective PAR2F240S agonists.

By introducing the F240S mutation into rat PAR2, we observed shifts in agonist potencies that mirrored the human PAR2F240S, suggesting that Phe240 is involved in determining agonist specificity of PAR2. Finally, differences in receptor signaling were paralleled in a cell growth assay. We suggest that the distinct pharmacological profile induced by this polymorphism will have important implications for the design of PAR-targeted agonists/antagonists and may contribute to, or be predictive of, an inflammatory disease.

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Published date: 15 December 2000

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Local EPrints ID: 166439
URI: http://eprints.soton.ac.uk/id/eprint/166439
ISSN: 0021-9258
PURE UUID: a76ecca7-eba1-422d-b0e2-9f4b88bd4a2c

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Date deposited: 28 Oct 2010 14:46
Last modified: 16 Jul 2019 23:50

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Contributors

Author: Steven J. Compton
Author: Jennifer A. Cairns
Author: Karan-Jane Palmer
Author: Bahjat AL-Ani
Author: Morley D. Hollenberg
Author: Andrew F. Walls

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