Department of Biology, University of York, York, UK.
Traffic. 2014 Jan;15(1):12-21. doi: 10.1111/tra.12128. Epub 2013 Oct 31.
Vesicle transport sorts proteins between compartments and is thereby responsible for generating the non-uniform protein distribution along the eukaryotic secretory and endocytic pathways. The mechanistic details of specific vesicle targeting are not yet well characterized at the molecular level. We have developed a cell-free assay that reconstitutes vesicle targeting utilizing the recycling of resident enzymes within the Golgi apparatus. The assay has physiological properties, and could be used to show that the two lobes of the conserved oligomeric Golgi tethering complex play antagonistic roles in trans-Golgi vesicle targeting. Moreover, we can show that the assay is sensitive to several different congenital defects that disrupt Golgi function and therefore cause glycosylation disorders. Consequently, this assay will allow mechanistic insight into the targeting step of vesicle transport at the Golgi, and could also be useful for characterizing some novel cases of congenital glycosylation disorders.
囊泡运输在隔室之间分拣蛋白质，从而负责在真核分泌和内吞途径中产生非均匀的蛋白质分布。特定囊泡靶向的机制细节在分子水平上尚未很好地表征。我们开发了一种无细胞测定法，该测定法利用高尔基体中驻留酶的再循环来重建囊泡靶向。该测定法具有生理特性，可用于表明保守寡聚高尔基体连接复合物的两个叶在跨高尔基囊泡靶向中发挥拮抗作用。此外，我们可以表明，该测定法对几种不同的先天性缺陷敏感，这些缺陷会破坏高尔基体的功能，从而导致糖基化紊乱。因此，该测定法将允许深入了解高尔基体中囊泡运输的靶向步骤，并且对于表征某些新型先天性糖基化疾病也可能有用。
