دورية
Cytosol-to-lysosome Transport of Free Polymannose-type Oligosaccharides
العنوان: | Cytosol-to-lysosome Transport of Free Polymannose-type Oligosaccharides |
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المؤلفون: | Saint-Pol, Agnès, Codogno, Patrice, Moore, Stuart E.H. |
المصدر: | Journal of Biological Chemistry; May 1999, Vol. 274 Issue: 19 p13547-13555, 9p |
مستخلص: | In hepatocellular carcinoma HepG2 cells, free polymannose-type oligosaccharides appearing in the cytosol during the biosynthesis and quality control of glycoproteins are rapidly translocated into lysosomes by an as yet poorly defined process (Saint-Pol, A., Bauvy, C., Codogno, P., and Moore, S. E. H. (1997)J. Cell Biol.136, 45–59). Here, we demonstrate an ATP-dependent association of [2-3H]mannose-labeled Man5GlcNAc with isolated rat liver lysosomes. This association was only observed in the presence of swainsonine, a mannosidase inhibitor, which was required for the protection of sedimentable, but not nonsedimentable, Man5GlcNAc from degradation, indicating that oligosaccharides were transported into lysosomes. Saturable high affinity transport (Kuptake, 22.3 μm, Vmax, 7.1 fmol/min/unit of β-hexosaminidase) was dependent upon the hydrolysis of ATP but independent of vacuolar H+/ATPase activity. Transport was inhibited strongly by NEM and weakly by vanadate but not by sodium azide, and, in addition, the sugar transport inhibitors phloretin, phloridzin, and cytochalasin B were without effect on transport. Oligosaccharide import did not show absolute specificity but was selective toward partially demannosylated and dephosphorylated oligosaccharides, and, furthermore, inhibition studies revealed that the free reducing GlcNAc residue of the oligosaccharide was of critical importance for its interaction with the transporter. These results demonstrate the presence of a novel lysosomal free oligosaccharide transporter that must work in concert with cytosolic hydrolases in order to clear the cytosol of endoplasmic reticulum-generated free oligosaccharides. |
قاعدة البيانات: | Supplemental Index |
تدمد: | 00219258 1083351X |
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DOI: | 10.1074/jbc.274.19.13547 |