Reference - Detail
|Author||Shirakawa R, Higashi T, Tabuchi A, Yoshioka A, Nishioka H, Fukuda M, Kita T, Horiuchi H.|
|Title||Munc13-4 is a GTP-Rab27-binding protein regulating dense core granule secretion in platelets.|
|Journal||J Biol Chem|
Platelets store self-agonists such as ADP and serotonin in dense core granules. Although exocytosis of these granules is crucial for hemostasis and thrombosis, the underlying mechanism is not fully understood. Here, we show that incubation of permeabilized platelets with unprenylated active mutant Rab27A-Q78L, wild type Rab27A, and Rab27B inhibited the secretion, whereas inactive mutant Rab27A-T23N and other GTPases had no effects. Furthermore, we affinity-purified a GTP-Rab27A-binding protein in platelets and identified it as Munc13-4, a homologue of Munc13-1 known as a priming factor for neurotransmitter release. Recombinant Munc13-4 directly bound to GTP-Rab27A and -Rab27B in vitro, but not other GTPases, and enhanced secretion in an in vitro assay. The inhibition of secretion by unprenylated Rab27A was rescued by the addition of Munc13-4, suggesting that Munc13-4 mediates the function of GTP-Rab27. Thus, Rab27 regulates the dense core granule secretion in platelets by employing its binding protein, Munc13-4.
|MeSH||Adenosine Diphosphate / metabolism Amino Acid Sequence Biological Transport Blood Platelets / metabolism Brain / metabolism Centrifugation, Density Gradient Cloning, Molecular Cytosol / metabolism DNA, Complementary / metabolism Dose-Response Relationship, Drug Glutathione Transferase / metabolism Guanosine 5'-O-(3-Thiotriphosphate) / metabolism Guanosine Triphosphate / metabolism* HeLa Cells Humans Models, Genetic Molecular Sequence Data Mutation Protein Binding Protein Structure, Tertiary Proteins / metabolism Proteins / physiology* Secretory Vesicles / metabolism* Time Factors rab GTP-Binding Proteins / metabolism* rab27 GTP-Binding Proteins|
|DNA material||pGEX-4T-3-Slac2-b-SHD (RDB17372)|