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

FEBS Letters

Djiane, Jean²  Devauchelle, Gérard¹  Cerutti, Martine¹  Cahoreau, Claire¹  Garnier, Laurence¹ 

[1] Laboratoire de Pathologie Comparée, Unité de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique, CNRS UA 1184, 30380 St-Christol-Lez-Alès, France;Unité d'Endocrinologie Moléculaire, Institut National de la Recherche Agronomique, 78352 Jouy-en-Josas, France

关键词: Baculovirus;    Cytoplasmic receptor domain;    Prolactin;    N-Glycosylation;    Ubiquitination;    PRL;    prolactin;    GH;    growth hormones;    SDS;    sodium dodecly sulfate;    PAGE;    polyacrylamide gel electrophoresis;    Sf;    Spodoptera frugiperda;    AcMNPV;    Autographa californica nuclear polyhedrosis virus;   

DOI  :  10.1016/0014-5793(94)00772-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 ≈ 9 kDa of the extracellular domain could be attributed to the N-glycosylation and another additional ≈ 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.

【 授权许可】

Unknown