Protein kinase C mediates platelet secretion and thrombus formation through protein kinase D2

• Published in: Blood Vol. 118; no. 2; pp. 416 - 424
• Main Authors: Konopatskaya, Olga, Matthews, Sharon A., Harper, Matthew T., Gilio, Karen, Cosemans, Judith M.E.M., Williams, Christopher M., Navarro, Maria N., Carter, Deborah A., Heemskerk, Johan W.M., Leitges, Michael, Cantrell, Doreen, Poole, Alastair W.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 14.07.2011; Americain Society of Hematology
• Subjects: Animals; Biological and medical sciences; Blood Platelets - drug effects; Blood Platelets - enzymology; Blood Platelets - metabolism; Female; Gene Expression Regulation, Enzymologic; Hematologic and hematopoietic diseases; Humans; Indoles - pharmacology; Male; Maleimides - pharmacology; Medical sciences; Mice; Mice, Knockout; Platelet Activation - drug effects; Platelet Activation - genetics; Platelet Aggregation - drug effects; Platelet Aggregation - genetics; Platelet Aggregation - physiology; Protein Kinase C - antagonists & inhibitors; Protein Kinase C - genetics; Protein Kinase C - metabolism; Protein Kinase C - physiology; Protein Kinase D2; Protein Kinase Inhibitors - pharmacology; Protein Kinases - genetics; Protein Kinases - metabolism; Protein Kinases - physiology; Secretory Vesicles - drug effects; Secretory Vesicles - metabolism; Signal Transduction - drug effects; Thrombosis - genetics; Thrombosis - metabolism; Protein kinase C; Platelet; Hematology; Enzyme; Secretion; Transferases; Non-specific serine/threonine protein kinase; Thrombus
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2010-10-312199

Platelets are highly specialized blood cells critically involved in hemostasis and thrombosis. Members of the protein kinase C (PKC) family have established roles in regulating platelet function and thrombosis, but the molecular mechanisms are not clearly understood. In particular, the conventional PKC isoform, PKCα, is a major regulator of platelet granule secretion, but the molecular pathway from PKCα to secretion is not defined. Protein kinase D (PKD) is a family of 3 kinases activated by PKC, which may represent a step in the PKC signaling pathway to secretion. In the present study, we show that PKD2 is the sole PKD member regulated downstream of PKC in platelets, and that the conventional, but not novel, PKC isoforms provide the upstream signal. Platelets from a gene knock-in mouse in which 2 key phosphorylation sites in PKD2 have been mutated (Ser707Ala/Ser711Ala) show a significant reduction in agonist-induced dense granule secretion, but not in α-granule secretion. This deficiency in dense granule release was responsible for a reduced platelet aggregation and a marked reduction in thrombus formation. Our results show that in the molecular pathway to secretion, PKD2 is a key component of the PKC-mediated pathway to platelet activation and thrombus formation through its selective regulation of dense granule secretion.