Glycoprotein 330 (gp330) is a member of a family of receptors related to the low density lipoprotein receptor (LDLR). Although several ligands have been shown to bind gp330 in solid-phase assays, the ability of gp330 to mediate ligand endocytosis has not been demonstrated. To develop a cellular model for gp330 function we screened a variety of cultured cell lines and identified several that expressed this protein, including immortalized rat type II pneumocytes and a human and two rodent tumor cell lines. Using type II pneumocytes, endocytosis of a previously described gp330 ligand, urokinase (uPA) complexed with plasminogen activator inhibitor-1 (uPA:PAI-1) and two new ligands, PAI-1 and pro-uPA, was demonstrated. RAP, the 39 kDa receptor-associated protein known to antagonize ligand binding to gp330 in solid-phase binding assays, completely inhibited both internalization and degradation of the radiolabeled ligands by type II pneumocytes. This suggested that the clearance of these ligands was dependent on either gp330 or the LDLR-related protein (LRP), which shares several ligand-binding characteristics with gp330. By using polyclonal antibodies to gp330, the cellular internalization and degradation of the ligands were inhibited by 30–50%; remaining ligand internalization and degradation activity could be partially inhibited by polyclonal antibodies against LRP. These findings indicate that gp330, like other LDLR family members, mediates endocytosis of its ligands. In addition, gp330 acts in concert with LRP in type II pneumocytes to mediate clearance of a variety of proteins involved in plasminogen activation, including uPA:PAI-1 complexes PAI-1 and pro-uPA.(ABSTRACT TRUNCATED AT 250 WORDS)

REFERENCES

Ashcom
J. D.
,
Tiller
S. E.
,
Dickerson
K.
,
Cravens
J. L.
,
Argraves
W. S.
,
Strickland
D. K.
(
1990
).
The human2-macroglobulin receptor: Identification of a 420-kD cell surface glycoprotein specific for the activated conformation of 2-macroglobulin.
J. Cell Biol
110
,
1041
1048
Barrett
A. J.
(
1981
).
2-Macroglobulin.
Meth. Enzymol
80
,
737
754
Beisiegel
U.
,
Schneider
W. J.
,
Goldstein
J. L.
,
Anderson
R. G.
,
Brown
M. S.
(
1981
).
Monoclonal antibodies to the low density lipoprotein receptor as probes for study of receptor-mediated endocytosis and the genetics of familial hypercholesterolemia.
J. Biol. Chem
256
,
11923
11931
Bertozzi
P.
,
Astedt
B.
,
Zenzius
L.
,
Lynch
K.
,
LeMaire
F.
,
Zapol
W.
,
Chapman
H. A.
(
1990
).
Depressed bronchioalveolar urokinase activity in patients with adult respiratory distress syndrome.
New Eng. J. Med
322
,
890
897
Chappell
D. A.
,
Fry
G. L.
,
Waknitz
M. A.
,
Muhonen
L. E.
,
Pladet
M. W.
,
Iverius
P.-H.
,
Strickland
D. K.
(
1993
).
Lipoprotein lipase induces catabolism of normal triglyceride rich lipoproteins via the low density receptor-related protein/2-macroglobulin receptor in vitro. A process mediated by cell surface proteoglycans.
J. Biol. Chem
268
,
14168
14175
Chatelet
F.
,
Brianti
E.
,
Ronco
P.
,
Roland
J.
,
Verroust
P.
(
1986
).
Ultrastructural localization by monoclonal antibodies of brush border antigens expressed by glomeruli. II. Extrarenal distribution.
Am. J. Pathol
122
,
512
519
Chen
W.
,
Goldstein
J. L.
,
Brown
M. S.
(
1990
).
NPXY, a sequence often found in cytoplasmic tails, is requied for coated pit-mediated internalization of the low density lipoprotein receptor.
J. Biol. Chem
265
,
3116
3123
Christensen
E. I.
,
Gliemann
J.
,
Moestrup
S. K.
(
1992
).
Renal tubule gp330 is a calcium binding receptor for endocytic uptake of protein.
J. Histochem. Cytochem
40
,
1481
1492
Cubellis
M. V.
,
Wun
T. C.
,
Blasi
F.
(
1990
).
Receptor-mediated internalization and degradation of urokinase is mediated by its specific inhibitor PAI-1.
EMBO J
9
,
1079
1085
Estreicher
A.
,
Mulhauser
J.
,
Carpentier
J.-L.
,
Orci
L.
,
Vassalli
J.-D.
(
1990
).
The receptor for urokinase type plasminogen activator polarizes expression of the protease to the leading edge of migrating monocytes and promotes the degradation of enzyme inhibitor complexes.
J. Cell Biol
111
,
783
792
Gafvels
M. E.
,
Caird
M.
,
Britt
D.
,
Jackson
C. L.
,
Patterson
D.
,
Strauss
J. F.
(
1993
).
Cloning of a cDNA encoding a putative human very low density lipoprotein/apolipoprotein E receptor and assignment of the gene to chromosome 9pter-p26.
Somat. Cell Mol. Genet
19
,
557
569
Gross
T. J.
,
Simon
R. H.
,
Sitrin
R. G.
(
1990
).
Expression of urokinase-type plasminogen activator by rat pulmonary alveolar epithelial cells.
Am. J. Respir. Cell Mol. Biol
3
,
449
456
Herz
J.
,
Hamann
U.
,
Rogne
S.
,
Myklebost
O.
,
Gausepohl
H.
,
Stanley
K. K.
(
1988
).
Surface location and high affinity for calcium of a 500 kd liver membrane protein closely related to the LDL-receptor suggest a physiological role as lipoprotein receptor.
EMBO J
7
,
4119
4127
Herz
J.
,
Kowal
R. C.
,
Ho
Y. K.
,
Brown
M. S.
,
Goldstein
J. L.
(
1990
).
Low density lipoprotein related-protein mediates endocytosis of monoclonal antibodies in cultured cells and rabbit liver.
J. Biol. Chem
265
,
21355
21362
Idell
S.
,
James
K. K.
,
Levin
E. G.
,
Schwartz
B. S.
,
Manchanda
N.
,
Maunder
R. J.
,
Martin
T. R.
,
McLarty
J.
,
Fair
D. S.
(
1990
).
Local abnormalities in coagulation and fibrinolytic pathways predispose to alveolar fibrin deposition in the adult respiratory distress syndrome.
J. Clin. Invest
84
,
695
705
Jensen
P. H.
,
Christensen
E. I.
,
Ebbesen
P.
,
Glieman
J.
,
Andersen
P. A.
(
1990
).
Lysosomal degradation of receptor-bound urokinase-type plasminogen activator is enhanced by its inhibitors in human trophoblastic choriocarinoma cells.
Cell Regul
1
,
1043
1056
Kerjaschki
D.
,
Farquhar
M. G.
(
1983
).
Immunocytochemical localization of the heyman nephritis antigen (gp330) in glomerular epithelial cells of normal Lewis rats.
J. Exp. Med
157
,
667
686
Kerjaschki
D.
,
Horvat
R.
,
Binder
S.
,
Susani
M.
,
Dekan
G.
,
Ojha
P. P.
,
Hillemanns
P.
,
Ulrich
W.
,
Donini
U.
(
1987
).
Identification of a 400-Kd protein in the brush borders of human kidney tubules that is similar to gp330, the nephritogenic antigen of rat Heymann Nephritis.
Am. J. Pathol
129
,
183
191
Korenberg
J. R.
,
Argraves
K. M.
,
Chen
X. N.
,
Tran
H.
,
Strickland
D. K.
,
Argraves
W. S.
(
1994
).
Chromosomal localization of human genes for the LDL receptor family member glycoprotein 330 (LRP2) and its associated protein RAP (LRPAP1).
Genomics
22
,
88
93
Kounnas
M. Z.
,
Argraves
W. S.
,
Strickland
D. K.
(
1992
).
The 39-kDa receptor-associated protein interacts with two members of the low density lipoprotein receptor family,2-macroglobulin receptor and glycoprotein 330.
J. Biol. Chem
267
,
21162
21168
Kounnas
M. Z.
,
Morris
R. E.
,
Thompson
M. R.
,
FitzGerald
D. J.
,
Strickland.
D. K.
,
Saelinger
C. B.
(
1992
).
The2-macroglobulin receptor/low density receptor-related protein binds and internalizes Pseudomonas exotoxin A.
J. Biol. Chem
267
,
12420
12423
Kounnas
M. Z.
,
Chappell
D. A.
,
Strickland
D. K.
,
Argraves
W. S.
(
1993
).
Glycoprotein 330, a member of the low density lipoprotein receptor family, binds lipoprotein lipase in vitro.
J. Biol. Chem
268
,
14176
14181
Kounnas
M. Z.
,
Henkin
J.
,
Argraves
W. S.
,
Strickland
D. K.
(
1993
).
Low density lipoprotein receptor-related protein/2-macroglobulin receptor mediates cellular uptake of pro-urokinase.
J. Biol. Chem
268
,
21862
21867
Kounnas
M. Z.
,
Haudenschild
C. C.
,
Strickland
D. K.
,
Argraves
W. S.
(
1994
).
Immunological localization of glycoprotein 330, low density lipoprotein receptor related protein and 39 kDa receptor associated protein in embryonic mouse tissues.
In Vivo
,
8
,
343
352
Kounnas
M. Z.
,
Stefansson
S.
,
Loukinova
E.
,
Argraves
K. M.
,
Strickland
D. K.
,
Argraves
W. S.
(
1994
).
Glycoprotein 330, an overview.
Ann. NY Acad. Sci
737
,
114
124
Lawrence
D.
,
Strandberg
L.
,
Grundstrom
T.
,
Ny
T.
(
1989
).
Purification of active human plasminogen activator inhibitor 1 from Escherichia coli.
J. Biochem
186
,
523
533
Mallampalli
R. K.
,
Floerchinger
C. S.
,
Hunninghake
G. W.
(
1992
).
Isolation and immortalization of rat pre-type II cell line.
In Vitro Cell. Dev. Biol
28
,
181
187
Marshall
B. C.
,
Sageser
D. S.
,
Rao
N. V.
,
Emi
M.
,
Hoidal
J. R.
(
1990
).
Alveolar epithelial cell plasminogen activator.
J. Biol. Chem
265
,
8198
8204
Marshall
B. C.
,
Xu
Q.-P.
,
Rao
N. V.
,
Brown
B.
,
Hoidal
J. R.
(
1992
).
Pulmonary epithelial cell urokinase-type plasminogen activator.
J. Biol. Chem
267
,
11462
11469
Moestrup
S. K.
,
Nielsen
S.
,
Andreasen
P.
,
Jorgensen
K. E.
,
Nykjaer
A.
,
Roigaard
H.
,
Gliemann
J.
,
Christensen
E. I.
(
1993
).
Epithelial glycoprotein-330 mediates endocytosis of plasminogen activator-plasminogen activator inhibitor type-1 complexes.
J. Biol. Chem
268
,
16564
16570
Moestrup
S. K.
(
1994
).
The2-macroglobulin receptor and epithelial glycoprotein-330: two giant receptors mediating endocytosis of multiple ligands.
Biochim. Biophys. Acta
1197
,
197
213
Munson
P. J.
,
Robard
D.
(
1980
).
Ligand: a versitile computerized approach for the characterization of ligand binding systems.
Anal. Biochem
107
,
220
239
Nykjaer
A.
,
Petersen
C. M.
,
Moller
B.
,
Jensen
P. H.
,
Moestrup
S. K.
,
Holtet
T. L.
,
Etzerodt
M.
,
Thogersen
H. C.
,
Munch
M.
,
Andreasen
P. A.
,
Gliemann
J.
(
1992
).
Purified2-macroglobulin receptor/LDL receptor-related protein binds urokinase plasminogen activator inhibitor type-1 complex: evidence that the 2-macroglobulin receptor mediates cellular degradation of urokinase receptor-bound complexes.
J. Biol. Chem
267
,
14543
14546
Nykjaer
A.
,
Kjoller
L.
,
Cohen
R.
,
Lawrence
D. A.
,
Garni-Wagner
B.-A.
,
Todd
R. F.
,
van Zonneveld
A.-J.
,
Gliemann
J.
,
Andreasen
P. A.
(
1994
).
Regions involved in binding of urokinase-type-1 inhibitor complex and pro-urokinase to the endocytic2-macroglobulin receptor/low density lipoprotein receptor related protein.
J. Biol. Chem
269
,
25668
25676
Orlando
R. A.
,
Kerjaschki
D.
,
Kurihara
H.
,
Biemesderfer
D.
,
Farquhar
M. G.
(
1992
).
gp330 associates with 44-kDa protein in the kidney to form the Heymann nephritis antigenic complex.
Proc. Nat. Acad. Sci. USA
89
,
6698
6702
Raychowdhury
R.
,
Niles
J. H.
,
McCluskey
R. T.
,
Smith
J. A.
(
1989
).
Autoimmune target in Heymann nephritis is a glycoprotein with homology to the LDL receptor.
Science
244
,
1163
1165
Saito
A.
,
Pietromonaco
S.
,
Loo
A. K.-C.
,
Farquhar
M. G.
(
1994
).
Complete cloning and sequencing of rat gp330/“megalin”, a distinctive member of the low density lipoprotein receptor family.
Proc. Nat. Acad. Sci. USA
91
,
9725
9729
Strickland
D. K.
,
Aschom
J. D.
,
William
S.
,
Burgess
W. H.
,
Migliorini
M.
,
Argraves
W. S.
(
1990
).
Sequence identity between the2-macroglobulin receptor and low density lipoprotein receptor-related protein suggest that this molecule is a multifunctional receptor.
J. Biol. Chem
265
,
17401
17404
Strickland
D. K.
,
Aschom
J. D.
,
William
S.
,
Battey
F.
,
Behre
E.
,
McTigue
K.
,
Battey
J. F.
,
Argraves
W. S.
(
1991
).
Primary structure of2-macroglobulin receptor-associated protein.
J. Biol. Chem
266
,
13364
13369
Williams
S.
,
Ashcom
J. D.
,
Argraves
W. S.
,
Strickland
D. K.
(
1992
).
A novel mechanism for controlling the activity of2-macroglobulin receptor/low density lipoprotein receptor-related protein.
J. Biol. Chem
267
,
9035
9040
Willnow
T. E.
,
Goldstein
J. L.
,
Orth
K.
,
Brown
M. S.
,
Herz
J.
(
1992
).
Low density lipoprotein receptor-related protein (LRP) and gp330 bind similar ligands including plasminogen activator/inhibitor complexes and lactoferrin, an inhibitor of chylomicron remenant clearance.
J. Biol. Chem
267
,
26172
26180
Zheng
G.
,
Bachinsky
D. R.
,
Stamenkovic
I.
,
Strickland
D. K.
,
Brown
D.
,
Andres
G.
,
McCluskey
R. T.
(
1994
).
Organ distribution in rats of two members of the low density lipoprotein receptor gene family, gp330 and LRP/2-macroglobulin receptor, and the receptor-associated protein (RAP).
J. Histochem. Cytochem
42
,
531
542
This content is only available via PDF.