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  • Title: Accumulation of UDP-GlcNAc into intracellular vesicles and occurrence of a carrier-mediated transport. Study with plasma-membrane-permeabilized mouse thymocytes.
    Author: Cecchelli R, Cacan R, Verbert A.
    Journal: Eur J Biochem; 1985 Nov 15; 153(1):111-6. PubMed ID: 2415359.
    Abstract:
    Metabolic labelling of mouse thymocytes with radioactive mannose or glucosamine leads to the formation of labelled GDP-Man and UDP-GlcNAc. Using isotonic ammonium chloride treatment which renders the plasma membrane of thymocytes permeable to sugar nucleotides, we demonstrate that, in contrast to GDP-Man, a pool of UDP-GlcNAc remains associated with the cells after plasma membrane permeabilization. These observations are confirmed in experiments in which permeabilized thymocytes are incubated with exogenous labelled GDP-Man and UDP-GlcNAc, and we show that only UDP-GlcNAc is accumulated into sealed intracellular vesicles. This accumulation is a saturable process which can be inhibited by UDP, demonstrating the occurrence of a specific carrier. This transport mechanism can be blocked by covalent attachment of a non-permeant inhibitor UDP-dialdehyde without affecting the N-acetylglucosaminyltransferase itself. The fact that this carrier-mediated transport is not inhibited by tunicamycin indicates that this translocation process of UDP-GlcNAc does not involve lipid intermediates.
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