PMID- 16965542
OWN - NLM
STAT- MEDLINE
DCOM- 20070108
LR  - 20230829
IS  - 1538-7933 (Print)
IS  - 1538-7836 (Linking)
VI  - 4
IP  - 12
DP  - 2006 Dec
TI  - PAR-1-dependent pp60src activation is dependent on protein kinase C and increased 
      [Ca2+]: evidence that pp60src does not regulate PAR-1-dependent Ca2+ entry in 
      human platelets.
PG  - 2695-703
AB  - BACKGROUND: The role of the tyrosine kinase pp60src in PAR-1-dependent Ca2+ entry 
      was investigated in human platelets. pp60src plays a role in thapsigargin 
      (TG)-evoked store-operated Ca2+ entry (SOCE), which is thought to be a major 
      component of thrombin-evoked Ca2+ entry. METHODS: pp60src tyr416 phosphorylation 
      was used to assess pp60src activation. Fura-2-loaded platelets were used to 
      monitor intracellular Ca2+ concentration ([Ca2+]i). RESULTS: Activation of PAR-1 
      with the specific agonist SFLLRN increased pp60src activation within 10 s. This 
      required phospholipase C (PLC) activity, Ca2+ release and a rise in intracellular 
      Ca2+. PP2, an inhibitor of Src-family tyrosine kinases, inhibited SFLLRN-evoked 
      Ca2+ entry, but also inhibited Ca2+ release and the extrusion of Ca2+ by the 
      plasma membrane Ca2+ ATPase. Actin polymerization and conventional protein kinase 
      C (cPKC) activity were required for TG- and SFLLRN-evoked pp60src activation. 
      Although Go6976, an inhibitor of cPKCs, inhibited TG-evoked SOCE, it had little 
      effect on SFLLRN- or thrombin-evoked Ca2+ entry. CONCLUSIONS: These data indicate 
      that stimulation of PAR-1 leads to activation of pp60src in human platelets, 
      through PLC and cPKC activation, Ca2+ release and actin polymerization. However, 
      as PKC and actin polymerization are not needed for SFLLRN-evoked Ca2+ entry, we 
      suggest that pp60src is also not required. The apparent inhibition of 
      SFLLRN-evoked Ca2+ entry by PP2 is likely to be secondary to reduced Ca2+ 
      release. These data argue against a contribution of this SOCE pathway to 
      PAR-1-dependent Ca2+ entry.
FAU - Harper, M T
AU  - Harper MT
AD  - Department of Physiology, Development and Neuroscience, University of Cambridge, 
      Cambridge, UK.
FAU - Sage, S O
AU  - Sage SO
LA  - eng
GR  - 064070/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060911
PL  - England
TA  - J Thromb Haemost
JT  - Journal of thrombosis and haemostasis : JTH
JID - 101170508
RN  - 0 (AG 1879)
RN  - 0 (Actins)
RN  - 0 (Carbazoles)
RN  - 0 (Chelating Agents)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Indoles)
RN  - 0 (Peptide Fragments)
RN  - 0 (Phospholipid Ethers)
RN  - 0 (Pyrimidines)
RN  - 0 (Receptor, PAR-1)
RN  - 0 (thrombin receptor peptide (42-47))
RN  - 136194-77-9 (Go 6976)
RN  - 139890-68-9 (1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid 
      acetoxymethyl ester)
RN  - 1Y6SNA8L5S (edelfosine)
RN  - 22144-77-0 (Cytochalasin D)
RN  - 526U7A2651 (Egtazic Acid)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 2.7.10.2 (Proto-Oncogene Proteins pp60(c-src))
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - EC 3.1.4.- (Type C Phospholipases)
RN  - EC 3.6.3.8 (Plasma Membrane Calcium-Transporting ATPases)
RN  - EC 3.6.3.8 (Sarcoplasmic Reticulum Calcium-Transporting ATPases)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Actins/metabolism
MH  - Blood Platelets/drug effects/*metabolism
MH  - Calcium/metabolism
MH  - *Calcium Signaling/drug effects
MH  - Carbazoles/pharmacology
MH  - Chelating Agents/pharmacology
MH  - Cytochalasin D/pharmacology
MH  - Cytoskeleton/drug effects/metabolism
MH  - Egtazic Acid/analogs & derivatives/pharmacology
MH  - Enzyme Activation/drug effects
MH  - Enzyme Inhibitors/pharmacology
MH  - Humans
MH  - In Vitro Techniques
MH  - Indoles/pharmacology
MH  - Peptide Fragments/pharmacology
MH  - Phospholipid Ethers/pharmacology
MH  - Phosphorylation
MH  - Plasma Membrane Calcium-Transporting ATPases/antagonists & inhibitors/metabolism
MH  - Protein Kinase C/antagonists & inhibitors/*metabolism
MH  - Proto-Oncogene Proteins pp60(c-src)/antagonists & inhibitors/*metabolism
MH  - Pyrimidines/pharmacology
MH  - Receptor, PAR-1/agonists/*metabolism
MH  - Sarcoplasmic Reticulum Calcium-Transporting ATPases/antagonists & 
      inhibitors/metabolism
MH  - Thapsigargin/pharmacology
MH  - Time Factors
MH  - Type C Phospholipases/antagonists & inhibitors/metabolism
EDAT- 2006/09/13 09:00
MHDA- 2007/01/09 09:00
CRDT- 2006/09/13 09:00
PHST- 2006/09/13 09:00 [pubmed]
PHST- 2007/01/09 09:00 [medline]
PHST- 2006/09/13 09:00 [entrez]
AID - S1538-7836(22)11795-2 [pii]
AID - 10.1111/j.1538-7836.2006.02207.x [doi]
PST - ppublish
SO  - J Thromb Haemost. 2006 Dec;4(12):2695-703. doi: 10.1111/j.1538-7836.2006.02207.x. 
      Epub 2006 Sep 11.