LDL receptor-related protein internalizes and degrades uPA-PAI-1 complexes and is essential for embryo implantation

doi:10.1016/0092-8674(92)90511-A

Cell

Volume 71, Issue 3, 30 October 1992, Pages 411-421

https://doi.org/10.1016/0092-8674(92)90511-A Get rights and content

Abstract

The low density lipoprotein receptor-related protein (LRP) is a large multifunctional clearance receptor that has been implicated in the hepatic uptake of chylomicron remnants and in the removal of protease-inhibitor complexes from the circulation and from the extracellular space. Disruption of the LRP gene in mice blocks development of LRP^−/− embryos around the implantation stage. The expression pattern of LRP in the postimplantation stage embryo is identical to that of urokinase, a plasminogen activator that confers invasive properties to migrating cells. We demonstrate that LRP mediates uptake and degradation of urokinase-type plasminogen activator-plasminogen activator inhibitor 1 complexes and propose that the inability of the giant cells to remove the inactive protease complexes from their surfaces interferes with implantation of the embryo.

References (55)

E. Appella et al.
The receptor-binding sequence of urokinase
J. Biol. Chem.
(1987)
W.-J. Chen et al.
NPXY, a sequence often found in cytoplasmic tails, is required for coated pitmediated internalization of the low density lipoprotein receptor
J. Biol. Chem.
(1990)
P.J. Fraker et al.
Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3α,6α-diphenylglycoluril
Biochem. Biophys. Res. Commun.
(1978)
J.L. Goldstein et al.
Receptormediated endocytosis of LDL in cultured cells
Meth. Enzymol.
(1983)
C.M. Hekman et al.
Endothelial cells produce a latent inhibitor of plasminogen activators that can be activated by denaturants
J. Biol. Chem.
(1985)
J. Herz et al.
Low density lipoprotein receptor-related protein mediates endocytosis of monoclonal antibodies in cultured cells and rabbit liver
J. Biol. Chem.
(1990)
J. Herz et al.
39-kDa protein modulates binding of ligands to low density lipoprotein receptor-related $protein α_{2} - macroglobulin$ receptor
J. Biol. Chem.
(1991)
C. Hilliker et al.
Assignment of the gene coding for the α₂-macroglobulin receptor to mouse chromosome 15 and to human chromosome 12q13-q14 by isotopic and nonisotopic in situ hybridization
Genomics
(1992)
P.H. Jensen et al.
Receptors for α₂-macroglobulin and pregnancy zone protein-proteinase complexes in the human placental syncytiotrophoblast
Placenta
(1988)
R.C. Kowal et al.
Opposing effects of apolipoproteins E and C on lipoprotein binding to low density lipoprotein receptor-related protein
J. Biol. Chem.
(1990)

T. Kristensen et al.

Evidence that the newly cloned low-density-lipoprotein receptor related protein (LRP) is the α₂-macroglobulin receptor

FEBS Lett.

(1990)

O.H. Lowry et al.

Protein measurement with the folin phenol reagent

J. Biol. Chem.

(1951)

A.P. McMahon et al.

The Wnt-1 (int-1) protooncogene is required for development of a large region of the mouse brain

Cell

(1990)

S.K. Moestrup et al.

Analysis of ligand recognition by the purified α₂-macroglobulin receptor (low density lipoprotein receptor-related protein)

J. Biol. Chem.

(1991)

O. Myklebost et al.

The gene for the human putative apo E receptor is on chromosome 12 in the segment q13–14

Genomics

(1989)

A. Nykjaer et al.

Purified α₂-macroglobulin receptor/LDL receptor-related protein binds urokinase plasminogen activator inhibitor type-1 complex: evidence that the α₂-macroglobulin receptor mediates cellular degradation of urokinase receptor-bound complexes

J. Biol. Chem.

(1992)

D. Olson et al.

Internalization of the urokinase-plasminogen activator inhibitor type-1 complex is mediated by the urokinase receptor

J. Biol. Chem.

(1992)

D.W. Russell et al.

Domain map of the LDL receptor: sequence homology with the epidermal growth factor precursor

Cell

(1984)

M.P. Stoppelli et al.

Autocrine saturation of pro-urokinase receptors on human A431 cells

Cell

(1986)

D.K. Strickland et al.

Sequence identity between the α₂-macroglobulin receptor and low density lipoprotein receptor-related protein suggests that this molecule is a multifunctional receptor

J. Biol. Chem.

(1990)

S. Strickland et al.

Plasminogen activator in early embryogenesis: enzyme production by trophoblast and parietal endoderm

Cell

(1976)

K.R. Thomas et al.

Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells

Cell

(1987)

J.A. van Mourik et al.

Purification of an inhibitor of plasminogen activator (anti-activator) synthesized by endothelial cells

J. Biol. Chem.

(1984)

O. Behrendtsen et al.

Metalloproteinases mediate extracellular matrix degradation by cells from mouse blastocyst outgrowths

Development

(1992)

U. Beisiegel et al.

Lipoprotein lipase enhances the binding of chylomicrons to low density lipoprotein receptor-related protein

A. Bradley

Production and analysis of chimeric mice

M.S. Brown et al.

The low-density lipoprotein receptor-related protein: double agent or decoy?

Curr. Opin. Lipid.

(1991)

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Cell

ArticleLDL receptor-related protein internalizes and degrades uPA-PAI-1 complexes and is essential for embryo implantation

Abstract

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Meth. Enzymol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Genomics

Placenta

J. Biol. Chem.

FEBS Lett.

J. Biol. Chem.

Cell

J. Biol. Chem.

Genomics

J. Biol. Chem.

J. Biol. Chem.

Cell

Cell

J. Biol. Chem.

Cell

Cell

J. Biol. Chem.

Metalloproteinases mediate extracellular matrix degradation by cells from mouse blastocyst outgrowths

Development

Lipoprotein lipase enhances the binding of chylomicrons to low density lipoprotein receptor-related protein

Production and analysis of chimeric mice

The low-density lipoprotein receptor-related protein: double agent or decoy?

Curr. Opin. Lipid.

Article
LDL receptor-related protein internalizes and degrades uPA-PAI-1 complexes and is essential for embryo implantation