PMID- 14581479
OWN - NLM
STAT- MEDLINE
DCOM- 20040212
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 279
IP  - 3
DP  - 2004 Jan 16
TI  - Hydrogen peroxide generation induces pp60src activation in human platelets: 
      evidence for the involvement of this pathway in store-mediated calcium entry.
PG  - 1665-75
AB  - Reactive oxygen species, such as H2O2, have been recognized as intracellular 
      messengers involved in several cell functions. Here we report the activation of 
      the tyrosine kinase pp60(src) by H2O2, a mechanism required for the activation of 
      store-mediated Ca2+ entry (SMCE) in human platelets. Treatment of platelets with 
      H2O2 resulted in a time- and concentration-dependent activation of pp60(src). 
      Incubation with GF 109203X, a protein kinase C (PKC) inhibitor, prevented 
      H2O2-induced pp60(src) activation. In contrast, dimethyl-BAPTA loading did not 
      affect this response, suggesting that activation of pp60(src) by H2O2 is 
      independent of increases in [Ca2+](i). Cytochalasin D, an inhibitor of actin 
      polymerization, significantly reduced H2O2-induced pp60(src) activation. We found 
      that platelet stimulation with thapsigargin (TG) plus ionomycin (Iono) or 
      thrombin induced rapid H2O2 production, a mechanism independent of elevations in 
      [Ca2+](i). Treatment of platelets with catalase attenuated TG plus Iono- and 
      thrombin-induced activation of pp60(src). In addition, catalase as well as the 
      pp60(src) inhibitor, PP1, reduced both the activation of SMCE and the coupling 
      between the hTrp1 and the IP(3)R type II without having any effect on the 
      maintenance of SMCE. Consistent with the role of PKC in the activation of 
      pp60(src) by H2O2, the PKC inhibitors GF 109202X and Ro-31-8220 were found to 
      reduced SMCE in platelets. This study suggests that platelet activation with TG 
      plus Iono or thrombin is associated with H2O2 production, which acts as a second 
      messenger by stimulating pp60(src) by a PKC-dependent pathway and is involved in 
      the activation of SMCE in these cells.
FAU - Rosado, Juan A
AU  - Rosado JA
AD  - Department of Physiology, Faculty of Veterinary Sciences, University of 
      Extremadura, Caceres 10071, Spain. jarosado@unex.es
FAU - Redondo, Pedro C
AU  - Redondo PC
FAU - Salido, Gines M
AU  - Salido GM
FAU - Gomez-Arteta, Emilio
AU  - Gomez-Arteta E
FAU - Sage, Stewart O
AU  - Sage SO
FAU - Pariente, Jose A
AU  - Pariente JA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20031027
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (4-amino-5-(4-methylphenyl)-7-(tert-butyl)pyrazolo(3,4-d)pyrimidine)
RN  - 0 (Pyrazoles)
RN  - 0 (Pyrimidines)
RN  - 0 (Reactive Oxygen Species)
RN  - 22144-77-0 (Cytochalasin D)
RN  - BBX060AN9V (Hydrogen Peroxide)
RN  - EC 1.11.1.6 (Catalase)
RN  - EC 2.7.10.2 (Proto-Oncogene Proteins pp60(c-src))
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - EC 3.4.21.5 (Thrombin)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Blood Platelets/*metabolism
MH  - Calcium/*metabolism
MH  - Catalase/pharmacology
MH  - Cytochalasin D/pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Enzyme Activation
MH  - Humans
MH  - Hydrogen Peroxide/metabolism/*pharmacology
MH  - Mice
MH  - Molecular Sequence Data
MH  - Protein Kinase C/physiology
MH  - Proto-Oncogene Proteins pp60(c-src)/*metabolism
MH  - Pyrazoles/pharmacology
MH  - Pyrimidines/pharmacology
MH  - Reactive Oxygen Species
MH  - Thrombin/pharmacology
EDAT- 2003/10/29 05:00
MHDA- 2004/02/13 05:00
CRDT- 2003/10/29 05:00
PHST- 2003/10/29 05:00 [pubmed]
PHST- 2004/02/13 05:00 [medline]
PHST- 2003/10/29 05:00 [entrez]
AID - S0021-9258(20)68722-1 [pii]
AID - 10.1074/jbc.M307963200 [doi]
PST - ppublish
SO  - J Biol Chem. 2004 Jan 16;279(3):1665-75. doi: 10.1074/jbc.M307963200. Epub 2003 
      Oct 27.