ERD2 proteins mediate ER retention of the HNEL signal of LRP's receptor-associated protein (RAP)

Guojun Bu, Stephanie Rennke, Hans J. Geuze

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

The 39 kDa receptor-associated protein (RAP) is a receptor antagonist that interacts with several members of the low density lipoprotein (LDL) receptor gene family. Upon binding to these receptors, RAP inhibits all ligand interactions with the receptors. Our recent studies have demonstrated that RAP is an endoplasmic reticulum (ER) resident protein and an intracellular chaperone for the LDL receptor-related protein (LRP). The HNEL sequence at the carboxyl terminus of RAP represents a novel ER retention signal that shares homology with the well-characterized KDEL signal. In the present study, using immunoelectron microscopy we demonstrate that cells stably transfected with human growth hormone (GH) tagged with either KDEL (GH+KDEL) or HNEL (GH+HNEL) signals exhibit ER and cis-Golgi localization typical of ER-retained proteins. Overexpression of not only GH+HNEL but also GH+KDEL cDNA in transfected cells results in saturation of ER retention receptors and secretion of endogenous RAP indicating that the two signals interact with the same ER retention receptor(s). The role of RAP in the maturation of LRP is further supported by the observation that functional LRP is reduced about 60% as a result of decreased intracellular RAP. Pulse-chase labeling and immunolocalization studies of ERD2.1 and ERD2.2 proteins in transfected cells demonstrate a long half-life and Golgi localization for both receptors. Finally, overexpression of either ERD2.1 or ERD2.2 proteins significantly increases the capacity of cells to retain both KDEL and HNEL-containing proteins. Taken together, our results thus demonstrate that ERD2 proteins are capable of retaining the novel ER retention signal associated with RAP.

Original languageEnglish (US)
Pages (from-to)65-73
Number of pages9
JournalJournal of cell science
Volume110
Issue number1
StatePublished - Jan 1 1997

Keywords

  • Chaperone
  • ER retention signal
  • ERD2 protein
  • Endocytic receptor

ASJC Scopus subject areas

  • Cell Biology

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