Pharmacological characterization of protease activated receptor-1 by a serum responsive element-dependent reporter gene assay: major role of calmodulin

Biochemical Pharmacology
Luc De VriesDidier Cussac

Abstract

We studied the protease activated receptor-1 coupling to a serum response element (SRE)-dependent luciferase activity readout in transfected COS-7 cells. Thrombin, with a pEC50 of 10.5, was 3000-fold more potent than the peptide agonists SFLLR and its derived compound C721-40 in stimulating luciferase activity, although the three agonists exhibited similar efficacy at the maximal concentration tested. Interestingly, SFLLR- and C721-40-induced luciferase activity was biphasic, suggesting that at least two populations of G proteins couple to the receptor. Further pharmacological characterization of this system was performed using selective protease activated receptor-1 antagonists. SCH203099 and ER-112787 blocked SFLLR-induced luciferase activity with similar potencies (pK(B) of 7), slightly higher than that exhibited by an arylisoxazole derivative compound from Merck (pK(B) of 6.1). These values correlated with their affinities established by competition binding experiments using [3H]-C721-40 as radioligand for protease activated receptor-1. Transduction mechanisms of protease activated receptor-1 coupling to SRE-dependent luciferase activity were examined using specific inhibitors. The Ca2+ chelator BAPTA-AM, as well as the cal...Continue Reading

References

Oct 1, 1992·Trends in Biochemical Sciences·R Treisman
Sep 29, 1995·The Journal of Biological Chemistry·J ZhangS E Rittenhouse
Apr 7, 1995·The Journal of Biological Chemistry·V Vouret-CraviariE Van Obberghen-Schilling
Apr 1, 1996·Circulation Research·T JiangS F Steinberg
Feb 14, 1997·The Journal of Biological Chemistry·M MolinoL F Brass
Jul 27, 1999·The Journal of Biological Chemistry·D WangJ M Quillan
Aug 18, 1999·Bioorganic & Medicinal Chemistry Letters·H S AhnY Wu
Nov 24, 1999·The Journal of Biological Chemistry·S P HeximerK J Blumer
Apr 15, 2000·Nature·M Nakanishi-MatsuiS R Coughlin
Jun 3, 2000·Annual Review of Pharmacology and Toxicology·L De VriesM G Farquhar
May 30, 2002·The Journal of Cell Biology·Olivier GenesteRichard Treisman
Dec 7, 2002·The Journal of Biological Chemistry·Ivana GaleticBrian A Hemmings
Jun 25, 2003·The Journal of Biological Chemistry·Jennifer S McCullarTheresa M Filtz
Oct 11, 2003·Arteriosclerosis, Thrombosis, and Vascular Biology·Takashi MinamiWilliam C Aird
Jun 24, 2004·The Journal of Pharmacology and Experimental Therapeutics·Nobuhisa TanakaYosuke Tojyo
Sep 17, 2004·Molecular Pharmacology·Susan F Steinberg
May 10, 2005·The Journal of Biological Chemistry·Joseph N McLaughlinHeidi E Hamm
Sep 28, 2005·The Journal of Biological Chemistry·Guoquan Liu, Tatyana A Voyno-Yasenetskaya

❮ Previous
Next ❯

Citations

Oct 2, 2009·Journal of Medicinal Chemistry·Michel PerezBruno Le Grand

❮ Previous
Next ❯

Related Concepts

Related Feeds

AKT Pathway

This feed focuses on the AKT serine/threonine kinase, which is an important signaling pathway involved in processes such as glucose metabolism and cell survival.

Calcium & Bioenergetics

Bioenergetic processes, including cellular respiration and photosynthesis, concern the transformation of energy by cells. Here is the latest research on the role of calcium in bioenergetics.