Evidence for N-glycosylation and ubiquitination of the prolactin receptor expressed in a baculovirus-insect cell system

FEBS Lett. 1994 Aug 22;350(2-3):230-4. doi: 10.1016/0014-5793(94)00772-1.

Abstract

The molecular mass of the rabbit prolactin receptor (rbPRLR) deduced from cDNA cloning is 66 kDa. However, the molecular mass of the full-length receptor expressed in the insect Sf9 cells was found to be 94 kDa. In order to explain this discrepancy, we analyzed the possible post-translational modifications of the PRLR. Sf9 cells were infected with recombinant baculoviruses in the presence of tunicamycin, an inhibitor of N-glycosylation. Results showed that an additional approximately 9 kDa of the extracellular domain could be attributed to the N-glycosylation and another additional approximately 20 kDa covalent modification occurred in the cytoplasmic part of the receptor. Western blot analysis, using anti-ubiquitin antibodies, revealed that the rbPRLR was ubiquitinated in its cytoplasmic domain.

MeSH terms

  • Animals
  • Baculoviridae
  • Glycosylation
  • In Vitro Techniques
  • Moths
  • Protein Processing, Post-Translational
  • Rabbits
  • Receptors, Prolactin / metabolism*
  • Recombinant Proteins / metabolism*
  • Ubiquitins / metabolism

Substances

  • Receptors, Prolactin
  • Recombinant Proteins
  • Ubiquitins