title: Munc13-4 is a GTP-Rab27-binding protein regulating dense core granule secretion in platelets.
abstract: 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.
authors.affiliation: Department of Geriatric Medicine, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Shirakawa
authors.forename: Tomohito
authors.initials: T
authors.lastname: Higashi
authors.forename: Arata
authors.initials: A
authors.lastname: Tabuchi
authors.forename: Akira
authors.initials: A
authors.lastname: Yoshioka
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nishioka
authors.forename: Mitsunori
authors.initials: M
authors.lastname: Fukuda
authors.forename: Toru
authors.initials: T
authors.lastname: Kita
authors.forename: Hisanori
authors.initials: H
authors.lastname: Horiuchi
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001692:Biological Transport; D001792:Blood Platelets / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005982:Glutathione Transferase / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000378:metabolism / Q000502:physiology*; D022142:Secretory Vesicles / Q000378:metabolism*; D013997:Time Factors; D020691:rab GTP-Binding Proteins / Q000378:metabolism*; D000075745:rab27 GTP-Binding Proteins
chemical_list: D018076:DNA, Complementary; D011506:Proteins; C430713:Unc13d protein, rat; D000075745:rab27 GTP-Binding Proteins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000244:Adenosine Diphosphate; D006160:Guanosine Triphosphate; D005982:Glutathione Transferase; C512956:Rab27b protein, rat; C066694:RAB27A protein, human; C512960:Rab27B protein, human; C512961:Rab27a protein, rat; D020691:rab GTP-Binding Proteins