Identification of the Minimal Functional Unit in the Low Density Lipoprotein Receptor-related Protein for Binding the Receptor-associated Protein (RAP)

Andersen, Olav M.; Christensen, Lisa Lystbæk; Christensen, Peter Bondo; Sørensen, Esben S.; Jacobsen, Christian; Moestrup, Søren K.; Etzerodt, Michael; Thøgersen, Henning

doi:10.1074/jbc.m000507200

Cited by 71 publications

(130 citation statements)

References 54 publications

(21 reference statements)

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“…The eluted product was concentrated and further purified using size-exclusion chromatography on either Superdex-75 or Superdex-200 (GE Healthcare) to isolate the monomeric forms in 20 mM HEPES, pH 7.4, 0.15 M NaCl, 0.005% Tween 20, and 5 mM CaCl 2 (HBS/Ca). LDLR CR cluster (CR.1-7) and its mutant were refolded as described (29). All proteins were verified by PAGE with GelCode Blue Safe (Thermo Scientific) staining and Western blotting with anti-myc mAb9E10.…”

Section: Methodsmentioning

confidence: 99%

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Mapping the Binding Region on the Low Density Lipoprotein Receptor for Blood Coagulation Factor VIII

Kurasawa

Shestopal

Karnaukhova

et al. 2013

Journal of Biological Chemistry

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Background: Low density lipoprotein receptor (LDLR) mediates clearance of blood coagulation factor VIII (FVIII). Results:The region of complement-type repeats 2-5 in LDLR was identified as the binding site for FVIII and for ␣-2-macroglobulin receptor-associated protein (RAP). Conclusion: Binding sites of LDLR for FVIII, and also for RAP, were characterized. Significance: This provides new data on LDLR structure and function and on FVIII catabolism.

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Section: Methodsmentioning

confidence: 99%

“…1 and 2). This strategy was assumed suitable, based on its previous use for mapping the RAP-binding site on LRP (29). The LDLR fragments were expressed in insect cells, which were previously shown capable of producing functionally active fragments of LRP and megalin (37,39,40).…”

Section: Expression Of Ldlr Fragments-mentioning

confidence: 99%

Mapping the Binding Region on the Low Density Lipoprotein Receptor for Blood Coagulation Factor VIII

Kurasawa

Shestopal

Karnaukhova

et al. 2013

Journal of Biological Chemistry

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“…These competition studies have now been extended to include the CD91-associated protein known as RAP. RAP is a well known CD91-binding protein that inhibits a number of CD91-mediated activities (23). Gp96-FITC (at a concentration, selected so as to be in the linear range of binding to RAW264.7 cells) was mixed with titrated molar excess amounts of RAP and incubated with paraformaldehyde-fixed RAW264.7 cells.…”

Section: Rap and Gp96mentioning

confidence: 99%

“…The reason for this observation is not entirely clear. RAP is known to bind to ligand-binding clusters II-IV of CD91, whereas ␣ 2 M is known to bind to clusters II and IV (14,23). The relative affinities of interactions of RAP and ␣ 2 M to CD91 have been reported to be similar in fluid phase studies in vitro.…”

Section: Rap Inhibits Re-presentation Of Gp96-chaperoned Peptides In mentioning

confidence: 99%

Essential role of CD91 in re-presentation of gp96-chaperoned peptides

Binder

Srivastava

2004

Proc. Natl. Acad. Sci. U.S.A.

185

152

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Heat shock proteins (HSPs) such as gp96 are released from cells as a result of necrotic cell death. The ability of endogenous HSPpeptide complexes to elicit antigen-specific T cells requires representation of the chaperoned peptides by antigen-presenting cells. Re-presentation requires the uptake of HSP-peptide complexes through a receptor, suggested to be the low-density lipoprotein receptor-related protein or CD91. We have used short interfering RNA for CD91 to show that, as antigen-presenting cells lose expression of CD91, their re-presenting ability undergoes a corresponding and dramatic decline. Furthermore, as the cells recover from extinction of CD91 expression, they regain the ability to re-present peptides. The ability of cells to bind ␣2 macroglobulin, a previously known CD91 ligand, or HSP gp96, and their ability to process peptides chaperoned by ␣2 macroglobulin, undergo identical variations. These results have been obtained from studies in vitro and from an assay that measures re-presentation in vivo. In additional studies in vivo, protective tumor immunity elicited by tumor-derived gp96 -peptide complexes is shown to be abrogated by anti-CD91 antisera. These studies show that CD91 is essential for re-presentation of gp96-chaperoned peptides by MHC molecules and have an important bearing on the mechanism of immunogenicity of necrotic cells.heat shock proteins ͉ tumor immunity ͉ ␣2-macroglobulin ͉ receptor-associated protein ͉ LRP H eat shock proteins (HSPs) purified from antigen-expressing cells chaperone antigenic peptides generated in that cell (1-4). Immunization with such HSP-peptide complexes purified from tumors or pathogen-infected cells elicits specific immunity directed against the tumor or pathogen, respectively (5-7). The mechanism of immunogenicity of HSP-peptide complexes is increasingly clear. The HSP-peptide complex is targeted to the antigen-presenting cells (APCs) through interaction of HSP with a receptor identified as CD91 (8,9). In case of the HSP gp96, it appears that the HSP-peptide complex is endocytosed into a compartment positive for Fc receptor and MHC I and negative for Rab5a, CD1, and transferrin (10). The subsequent fate of the HSP molecule itself remains unclear, but the peptide is transported to the cytosol through a mechanism yet to be determined. The peptide is further transported into the endoplasmic reticulum through a transporterassociated with antigen processing-dependent (9, 11) or -independent (see refs. 11 and 12) mechanism. Following a classical pathway within the endoplasmic reticulum, the peptide is charged onto a cognate MHC I molecule, and the MHC I-peptide complex present at the cell surface now stimulates cognate CD8 ϩ T cells. Because the endocytosed HSP-peptide complex is taken up into a compartment that is MHC I ϩ (10), the possibility that the gp96-chaperoned peptide is charged onto the MHC I in that compartment, independently of the endoplasmic reticulum, merits consideration.Among the several HSPs that follow this path, the endoplasmic reticulum-r...

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“…However, RAP binds with a much higher affinity to the cluster of 11 LA repeats than to the Vps10p domain. Recent studies on LRP have shown that high affinity RAP binding depends on pairs of LA repeats each harboring surface exposed acidic residues between Cys IV and Cys V of the ϳ40 residue repeats (29). SorLA has five LA repeats with exposed Asp/Glu residues (four in consecutive repeats) that may explain the RAP binding.…”

Section: Figmentioning

confidence: 99%

Activation and Functional Characterization of the Mosaic Receptor SorLA/LR11

Jacobsen

Madsen

Jacobsen

et al. 2001

Journal of Biological Chemistry

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159

190

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We previously isolated and sequenced the ϳ250-kDa type 1 receptor sorLA/LR11, a mosaic protein with elements characterizing the Vps10p domain receptor family as well as the low density lipoprotein receptor family. The N terminus of the Vps10p domain comprises a consensus sequence for cleavage by furin ( 50 RRKR 53 ) that precedes a truncation found in sorLA isolated from human brain. Here we show that sorLA, like sortilin-1/ neurotensin receptor-3, whose lumenal domain consists of a Vps10p domain only, is synthesized as a proreceptor that is cleaved by furin in late Golgi compartments. We show that the truncation conditions the Vps10p domain for propeptide inhibitable binding of neuropeptides and the receptor-associated protein. We further demonstrate that avid binding of the receptor-associated protein, apolipoprotein E, and lipoprotein lipase not inhibited by propeptide occurs to sites located in other lumenal domains. In transfected cells, about 10% of fulllength sorLA were expressed on the cell surface capable of mediating endocytosis. However, the major pool of receptors was found in late Golgi compartments, suggesting possible interaction with newly synthesized ligands. The results show that sorLA, following activation by truncation, binds multiple ligands and may mediate both endocytosis and sorting.

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Identification of the Minimal Functional Unit in the Low Density Lipoprotein Receptor-related Protein for Binding the Receptor-associated Protein (RAP)

Cited by 71 publications

References 54 publications

Mapping the Binding Region on the Low Density Lipoprotein Receptor for Blood Coagulation Factor VIII

Mapping the Binding Region on the Low Density Lipoprotein Receptor for Blood Coagulation Factor VIII

Essential role of CD91 in re-presentation of gp96-chaperoned peptides

Activation and Functional Characterization of the Mosaic Receptor SorLA/LR11

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