title: Fluorescent microscopy as a tool to elucidate dysfunction and mislocalization of Golgi glycosyltransferases in COG complex depleted mammalian cells.
abstract: Staining of molecules such as proteins and glycoconjugates allows for an analysis of their localization within the cell and provides insight into their functional status. Glycosyltransferases, a class of enzymes which are responsible for glycosylating host proteins, are mostly localized to the Golgi apparatus, and their localization is maintained in part by a protein vesicular tethering complex, the conserved oligomeric Golgi (COG) complex. Here we detail a combination of fluorescent lectin and immuno-staining in cells depleted of COG complex subunits to examine the status of Golgi glycosyltransferases. The combination of these techniques allows for a detailed characterization of the changes in function and localization of Golgi glycosyltransferases with respect to transient COG subunit depletion.
authors.affiliation: Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
authors.forename: Rose A
authors.initials: RA
authors.lastname: Willett
authors.forename: Irina D
authors.initials: ID
authors.lastname: Pokrovskaya
authors.forename: Vladimir V
authors.initials: VV
authors.lastname: Lupashin
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics*; D005455:Fluorescent Antibody Technique / Q000379:methods; D016695:Glycosyltransferases / Q000032:analysis* / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D034741:RNA, Small Interfering; D016695:Glycosyltransferases