Anaphylatoxin C3a induces rapid protein phosphorylation in guinea pig platelets.
We investigated C3a signal transduction using guinea pig platelets. Anaphylatoxin C3a induced cytosolic calcium influx and turnover of inositol phospholipids and caused protein phosphorylation via specific C3a receptors on guinea pig platelets. Phosphorylation of 40 kDa and 20 kDa proteins was detected within 30 s after C3a stimulation. From phosphoamino acid analysis of both proteins, it was found that about 80% was serine and 20% was threonine. Phosphorylation was decreased by pretreatment of platelets with calcium blockers, diltiazem and TMB-8. A protein kinase C inhibitor, staurosporine, effectively blocked phosphorylation of both proteins, but H-7 did not. The effects of these inhibitors on platelet aggregation were correlated with their effects on protein phosphorylation. These results suggest that protein-serine/threonine phosphorylation is important for C3a signal transduction in guinea pig platelets and that the isoenzyme of protein kinase C or another kinase probably participates in such protein phosphorylation. C3a also caused protein-tyrosine phosphorylation of 56 kDa and 33-36 kDa proteins within 30 s. Staurosporine effectively blocked phosphorylation of these bands. It is suggested that tyrosine kinase also may be involved in C3a signalling in guinea pig platelets.
Fukuoka
Y
Y
Department of Molecular Immunology, Tohoku University, Sendai, Japan.
Tachibana
T
T
Yasui
A
A
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D002118:Calcium / Q000097:blood; D015926:Complement C3a / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006801:Humans; D007295:Inositol Phosphates / Q000097:blood; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000276:immunology; D011493:Protein Kinase C / Q000097:blood / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000097:blood / Q000378:metabolism*; D015398:Signal Transduction
D007295:Inositol Phosphates; D015926:Complement C3a; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D002118:Calcium