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N-Glycosylation of membrane glycoproteins in retinol-deficient rat liver

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Abstract

The effect of vitamin A deficiency onN-linked oligosaccharides of membrane glycoproteins was studied in rat liver in order to evaluate the suggested role of retinol in proteinN-glycosylation. First, oligosaccharides of newly synthesized glycoproteins from rough endoplasmic reticulum of vitamin A deficient liver were compared with that of pair-fed controls. Oligosaccharides were metabolically labelled withd-[2-3H]mannose, released from the glycoproteins with endoglycosidase H, purified by reversed phase HPLC and ion exchange chromatography, and were reduced with sodium borohydride. HPLC fractionation of the oligosaccharide alditols showed that the glycoproteins carried mainly four oligosaccharide species, Glc1Man9GlcNAc2, Man9GlcNAc2, Man8GlcNAc2 and Man7GlcNAc2, in identical relative amounts in the vitamin A deficient and the control tissue. In particular, no increase in the proportion of short chain oligosaccharides was noted in vitamin A deficient liver. Second, the number ofN-linked oligosaccharides was estimated in dipeptidylpeptidase IV (DPP IV), a major glycoprotein constituent of the hepatic plasma membrane, comparing the newly synthesized glycoprotein from rough endoplasmic reticulum and the mature form of DPP IV from the plasma membrane. No evidence was obtained that retinol deficiency caused incomplete glycosylation of this membrane glycoprotein. From these data, the suggested role of retinol as a cofactor involved in the synthesis ofN-linked oligosaccharides of glycoproteins must be questioned.

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Abbreviations

DolP:

Dolichyl phosphate

DolPP:

dolichyl pyrophosphoryl

RetPMan:

retinyl phosphate mannose

DPP IV:

dipeptidyl peptidase IV (EC 3.4.14.5)

endo H:

endo-β-N-acetylglucosaminidase H (EC 3.2.1.96)

endo F:

endo-β-N-acetylglucosaminidase F (EC 3.2.1.96)

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. Institut für Klinische Chemie und Biochemie, Freie Universität Berlin, Spandauer Damm 130, D-1000, Berlin 19, Germany

    R. Tauber, E. Köttgen & Ch. Karasiewicz

  2. Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Arnimallee 22, D-1000, Berlin 33, Germany

    R. Nuck & W. Reutter

  3. Medizinische Universitätsklinik Freiburg, Hugstetter Straße 55, D-7800, Freiburg, Germany

    W. Gerok

  4. Medizinische Klinik III der Rheinisch-Westfälischen Technischen Hochschule, Pauwelsstraße, D-5100, Aachen, Germany

    R. Büchsel

  5. Universitäts-HNO-Klinik der Universität Freiburg, Kilianstraße, D-7800, Freiburg, Germany

    W. Lohle

Authors
  1. R. Tauber
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  2. R. Nuck
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  3. W. Gerok
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  4. R. Büchsel
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  5. E. Köttgen
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  6. W. Lohle
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  7. Ch. Karasiewicz
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  8. W. Reutter
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Tauber, R., Nuck, R., Gerok, W. et al. N-Glycosylation of membrane glycoproteins in retinol-deficient rat liver. Glycoconjugate J 9, 132–140 (1992). https://doi.org/10.1007/BF00780760

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  • DOI: https://doi.org/10.1007/BF00780760

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