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Dissecting Functions of the Conserved Oligomeric Golgi Tethering Complex Using a Cell‐Free Assay
Author(s) -
Cottam Nathanael P.,
Wilson Katherine M.,
Ng Bobby G.,
Körner Christian,
Freeze Hudson H.,
Ungar Daniel
Publication year - 2014
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/tra.12128
Subject(s) - golgi apparatus , biology , microbiology and biotechnology , endocytic cycle , copi , vesicle , secretory pathway , glycosylation , transport protein , vesicular transport protein , endosome , vesicular transport proteins , endoplasmic reticulum , cell , biochemistry , endocytosis , intracellular , vacuolar protein sorting , membrane
Intra‐Golgi vesicle transport was reconstituted in vitro using fluorescently marked galactosyltransferase as a marker. Microscopic analysis of transport reactions provides the readout, and shows physiological properties of the reconstitution, including protein, energy and temperature dependence. The new assay was used to show antagonistic functions of the two halves of the conserved oligomeric Golgi complex during trans ‐Golgi vesicle tethering. The assay will be particularly useful to study molecular determinants of tethering, and human congenital glycosylation disorders that affect Golgi trafficking.