Purification and functional analysis of a Rab27 effector munc 13-4 using a semi-intact platelet dense-granule secretion assay.
We have demonstrated that small GTPase Rab27 regulates dense-granule secretion in platelets. Using Rab27a affinity chromatography, we purified Munc 13-4 as a novel Rab27a interacting protein from platelet cytosol. This chapter describes the purification of Munc 13-4 and an in vitro assay system analyzing the mechanism of dense-granule secretion in platelets. The activity of Munc 13-4 is tested in this assay.
Ryutaro
R
Shirakawa
Tomohito
T
Higashi
Hirokazu
H
Kondo
Akira
A
Yoshioka
Toru
T
Kita
Hisanori
H
Horiuchi
D001792:Blood Platelets / Q000378:metabolism / Q000502:physiology*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000502:physiology*; D000075745:rab27 GTP-Binding Proteins
D008565:Membrane Proteins; D011994:Recombinant Proteins; C507969:UNC13D protein, human; D000075745:rab27 GTP-Binding Proteins; D012701:Serotonin; C066694:RAB27A protein, human; D020691:rab GTP-Binding Proteins