Glycoconjugate Journal

, Volume 9, Issue 3, pp 132–140 | Cite as

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

  • R. Tauber
  • R. Nuck
  • W. Gerok
  • R. Büchsel
  • E. Köttgen
  • W. Lohle
  • Ch. Karasiewicz
  • W. Reutter
Papers

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.

Keywords

Vitamin A retinol N-linked oligosaccharides membrane glycoproteins dipeptidylpeptidase IV 

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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Copyright information

© Chapman & Hall 1992

Authors and Affiliations

  • R. Tauber
    • 1
  • R. Nuck
    • 2
  • W. Gerok
    • 3
  • R. Büchsel
    • 4
  • E. Köttgen
    • 1
  • W. Lohle
    • 5
  • Ch. Karasiewicz
    • 1
  • W. Reutter
    • 2
  1. 1.Institut für Klinische Chemie und BiochemieFreie Universität BerlinBerlin 19Germany
  2. 2.Institut für Molekularbiologie und BiochemieFreie Universität BerlinBerlin 33Germany
  3. 3.Medizinische Universitätsklinik FreiburgFreiburgGermany
  4. 4.Medizinische Klinik III der Rheinisch-Westfälischen Technischen HochschuleAachenGermany
  5. 5.Universitäts-HNO-Klinik der Universität FreiburgFreiburgGermany

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