TY - JOUR
T1 - Purification and functional analysis of a Rab27 effector Munc13-4 Using a semiintact platelet dense-granule secretion assay
AU - Shirakawa, Ryutaro
AU - Higashi, Tomohito
AU - Kondo, Hirokazu
AU - Yoshioka, Akira
AU - Kita, Toru
AU - Horiuchi, Hisanori
N1 - Funding Information:
We thank Dr. Y. Nozawa for Rab27a cDNA, the Kyoto Red Cross Blood Center for providing outdated platelets, and T. Matsubara for excellent technical assistance. R.S. is a recipient of Fellowship of Japan Society for the Promotion of Science. This work was supported by the Japan Ministry of Education, Culture, Sports, Science, and Technology Research Grants 16–1244 (to R.S.), 15590740 (to H. H.), and 16209031 (to T. K.), by Health and Labour Sciences Research Grant H14‐Chouju‐012 from the Ministry of Health Labour and Welfare (to T. K. and H. H.), and in part by grants from the Takeda Science Foundation (to H.H).
PY - 2005
Y1 - 2005
N2 - We have demonstrated that small GTPase Rab27 regulates dense-granule secretion in platelets. Using Rab27a affinity chromatography, we purified Munc13-4 as a novel Rab27a interacting protein from platelet cytosol. This chapter describes the purification of Munc13-4 and an in vitro assay system analyzing the mechanism of dense-granule secretion in platelets. The activity of Munc13-4 is tested in this assay.
AB - We have demonstrated that small GTPase Rab27 regulates dense-granule secretion in platelets. Using Rab27a affinity chromatography, we purified Munc13-4 as a novel Rab27a interacting protein from platelet cytosol. This chapter describes the purification of Munc13-4 and an in vitro assay system analyzing the mechanism of dense-granule secretion in platelets. The activity of Munc13-4 is tested in this assay.
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U2 - 10.1016/S0076-6879(05)03067-3
DO - 10.1016/S0076-6879(05)03067-3
M3 - Review article
C2 - 16473638
AN - SCOPUS:32344448799
SN - 0076-6879
VL - 403
SP - 778
EP - 788
JO - Methods in Enzymology
JF - Methods in Enzymology
M1 - 67
ER -