Dissection of receptor folding and ligand-binding property with functional minireceptors of LDL receptor-related protein

Lynn M. Obermoeller-McCormick, Yonghe Li, Hiroshi Osaka, David J. FitzGerald, Alan L. Schwartz, Guojun D Bu

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

The LDL receptor-related protein (LRP) is a large, multifunctional endocytic receptor that binds and endocytoses a variety of structurally and functionally distinct ligands. LRP contains four putative ligand-binding domains. However, only domains II, III and IV, but not domain I, bind the receptor-associated protein (RAP), a molecular chaperone and universal antagonist for LRP. In order to dissect the function of RAP in LRP folding and to examine the ligand-binding properties of LRP, we generated LRP minireceptors that represent each of the four putative ligand-binding domains (termed mLRP1, mLRP2, mLRP3 and mLRP4, respectively). We found that proper folding and trafficking of mLRP2, mLRP3, mLRP4, but not mLRP1, is facilitated by coexpression of RAP. When these mLRPs were stably expressed in Chinese Hamster Ovary cells that lack the endogenous LRP, we found that each of these receptors was processed and traffics through the secretory pathway. Cell surface expression of these minireceptors was quantitatively examined by flow cytometric analyses. Using these minireceptor cell lines to map the ligand-binding domains, we found that although the majority of LRP ligands bind to both domain II and domain IV, Pseudomonas exotoxin A utilizes only domain IV for its binding to LRP. We conclude that while domains II and IV of LRP share many ligand-binding properties, each of the putative ligand-binding domains of LRP is unique in its contribution to ligand binding.

Original languageEnglish (US)
Pages (from-to)899-908
Number of pages10
JournalJournal of Cell Science
Volume114
Issue number5
StatePublished - 2001
Externally publishedYes

Fingerprint

Low Density Lipoprotein Receptor-Related Protein-1
Dissection
Ligands
Molecular Chaperones
Secretory Pathway
Protein Folding
Endocytosis
Cricetulus
Ovary
Proteins

Keywords

  • Endocytosis
  • LRP
  • Minireceptor
  • RAP
  • Receptor folding

ASJC Scopus subject areas

  • Cell Biology

Cite this

Obermoeller-McCormick, L. M., Li, Y., Osaka, H., FitzGerald, D. J., Schwartz, A. L., & Bu, G. D. (2001). Dissection of receptor folding and ligand-binding property with functional minireceptors of LDL receptor-related protein. Journal of Cell Science, 114(5), 899-908.

Dissection of receptor folding and ligand-binding property with functional minireceptors of LDL receptor-related protein. / Obermoeller-McCormick, Lynn M.; Li, Yonghe; Osaka, Hiroshi; FitzGerald, David J.; Schwartz, Alan L.; Bu, Guojun D.

In: Journal of Cell Science, Vol. 114, No. 5, 2001, p. 899-908.

Research output: Contribution to journalArticle

Obermoeller-McCormick, LM, Li, Y, Osaka, H, FitzGerald, DJ, Schwartz, AL & Bu, GD 2001, 'Dissection of receptor folding and ligand-binding property with functional minireceptors of LDL receptor-related protein', Journal of Cell Science, vol. 114, no. 5, pp. 899-908.
Obermoeller-McCormick, Lynn M. ; Li, Yonghe ; Osaka, Hiroshi ; FitzGerald, David J. ; Schwartz, Alan L. ; Bu, Guojun D. / Dissection of receptor folding and ligand-binding property with functional minireceptors of LDL receptor-related protein. In: Journal of Cell Science. 2001 ; Vol. 114, No. 5. pp. 899-908.
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