Molecular mechanism of thrombin receptor-specific 33 kDa protein kinase
| Project/Area Number |
13680711
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Functional biochemistry
|
|---|
| Research Institution | Mie University |
|---|
Principal Investigator |
IDO Masaru Mie University, Hospital, Associate Professor, 医学部附属病院, 助教授 (90167263)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Koji Mie University, Faculty of Medicine, Professor, 医学部, 教授 (70077808)
|
|---|
| Project Period (FY) |
2001 – 2002
|
| Project Status |
Completed (Fiscal Year 2002)
|
| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
|
|---|
| Keywords | Thrombin receptor / Serine / threonine kinase / Platelet / μ-calpain |
|---|
| Research Abstract |
Thrombin is a multifunctional serine protease that plays an important role in hemostasis and wound healing. Protease-activated receptor 1 (PAR1/ thrombin receptor) is a member of the G protein-coupled receptor (GPCR) containing seven transmembrane domains. Thrombin cleaves the receptor at the Arg41/Ser42 peptide bond and unmasks a tethered amino-terminal sequence beginning with the Ser-Phe-Leu-Leu-Arg-Asn (thrombin receptor agonist peptide/TRAP) ; subsequent binding of this ligand sequence to the body of the receptor is coupled with a transmembrane signaling mediated by G proteins. Phosphorylation of GPCR by G protein-coupled receptor kinases (GRKs) is thought to play a central role in receptor regulation. Recently, casein kinase I (CKI) was reported to phosphorylate GPCR. We previously identified a novel 33-kDa Ser/Thr protein kinase (PK33) in thrombin-stimulated platelets that participates in the phosphorylation of cytoplasmic tail of PAR-1. In the present study, we examined whether PK33 belongs to the same family of GRKs or CKI. PK33 was partially purified from thrombin-stimulated platelets through chromatography using HiTrap-DEAE, HiTrap-SP, HiTrap-Blue, and HiTrap-Heparin. Fractions after HiTrap-Heparin were examined by Western blotting using anti-GRK2, anti-GRK3, anti-GRK5 or anti-CKI antibody and in-gel renaturation protein kinase assay for PK33. GRK2, GRK3 and CKI were eluted in the HiTrap-Heparin-bound fractions differently from PK33. GRK5 did not bind to HiTrap-Heparin. The results of this study suggest that PK33 is a novel protein kinase distinct from GRK2, GRK3, GRK5 or CKI.
|
Report
(3 results)
Research Products
(13 results)
-
-
-
-
-
-
-
-
[Publications] Shimizu S, Gabazza EC, Taguchi O, Yasui H, Taguchi Y, Hayashi T, Ido M, Shimizu T, Nakagaki T, Kobayashi H, Fukudome K, Tsuneyoshi N, D'Alessandro-Gabazza CN, Izumizaki M, Iwase M, Homma I, Adachi Y and Suzuki K: "Activated Protein C Inhibits the Expression of Platelet-derived Growth Factor in the Lung"Am J Respir Crit Care Med. 167. 1416-1426 (2003)
Description
「研究成果報告書概要(欧文)」より
Related Report
-
-
-
-
-
