Sacher M, Jiang Y, Barrowman J, Scarpa A, Burston J, Zhang L, Schieltz D, Yates J R, Abeliovich H, Ferro-Novick S
Howard Hughes Medical Institute and the Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA.
EMBO J. 1998 May 1;17(9):2494-503. doi: 10.1093/emboj/17.9.2494.
We previously identified BET3 by its genetic interactions with BET1, a gene whose SNARE-like product acts in endoplasmic reticulum (ER)-to-Golgi transport. To gain insight into the function of Bet3p, we added three c-myc tags to its C-terminus and immunopurified this protein from a clarified detergent extract. Here we report that Bet3p is a member of a large complex ( approximately 800 kDa) that we call TRAPP (transport protein particle). We propose that TRAPP plays a key role in the targeting and/or fusion of ER-to-Golgi transport vesicles with their acceptor compartment. The localization of Bet3p to the cis-Golgi complex, as well as biochemical studies showing that Bet3p functions on this compartment, support this hypothesis. TRAPP contains at least nine other constituents, five of which have been identified and shown to be highly conserved novel proteins.
我们之前通过与BET1的遗传相互作用鉴定出BET3,BET1基因的类SNARE产物在内质网(ER)到高尔基体的转运过程中发挥作用。为深入了解Bet3p的功能,我们在其C末端添加了三个c-myc标签,并从澄清的去污剂提取物中免疫纯化了该蛋白。在此我们报告,Bet3p是一个大型复合物(约800 kDa)的成员,我们将其称为TRAPP(转运蛋白颗粒)。我们提出TRAPP在ER到高尔基体的转运囊泡与其受体区室的靶向和/或融合过程中起关键作用。Bet3p定位于顺式高尔基体复合物,以及生化研究表明Bet3p在该区室发挥功能,均支持这一假说。TRAPP至少还包含其他九种成分,其中五种已被鉴定并证明是高度保守的新蛋白。