We showed previously that a large chondroitin sulfate proteoglycan, PG-M (also known as versican), inhibits cell-substratum adhesion, while basement membrane heparan sulfate proteoglycan (recently named perlecan) does not (Yamagata et al. (1989) J. Biol. Chem. 264, 8012–8018). To extend our understanding of the adhesive function of these proteoglycans, we examined the pericellular localization of the proteoglycans and their ligands and also that of some matrix receptors and cytoskeletal molecules in various fibroblast culture systems. PG-M was abundant in the subcellular space of fibroblasts, but was excluded selectively from focal contacts where vinculin, integrins and fibronectin were localized. Hyaluronan, CD44 and tenascin were distributed similarly as PG-M. In contrast, perlecan was associated with fibronectin and was included in focal contacts. Syndecan-1, a membrane heparan sulfate/chondroitin sulfate proteoglycan, was associated with fibronectin at the cell surface, partly at focal contacts and in association with stress fibers. Thus, complexes of PG-M with hyaluronan, tenascin and CD44, are not involved in focal contacts. On the other hand, perlecan and syndecan-1 together with fibronectin may participate in focal contacts. The difference in localization between these proteoglycans may be related to their glycosaminoglycan content and to their distinctive roles in cell-substratum adhesion.

REFERENCES

Akiyama
S. K.
,
Yamada
S. S.
,
Yamada
K. M.
(
1986
).
Characterization of a 140kD avian cell surface antigen as a fibronectin-binding molecule.
J. Cell Biol
102
,
442
448
Aruffo
A.
,
Stamenkovic
I.
,
Melnick
M.
,
Underhill
C. B.
,
Seed
B.
(
1990
).
CD44 is the principal cell surface receptor for hyaluronate.
Cell
61
,
1303
1313
Avnur
Z.
,
Geiger
B.
(
1984
).
Immunocytochemical localization of native chondroitin-sulfate in tissues and cultured cells using specific monoclonal antibody.
Cell
38
,
811
822
Avnur
Z.
,
Geiger
B.
(
1985
).
Spatial interrelationships between proteoglycans and extracellular matrix proteins in cell cultures.
Exp. Cell Res
158
,
321
322
Bernfield
M.
,
Kokenyesi
R.
,
Kato
M.
,
Hinkes
M. T.
,
Spring
J.
,
Gallo
R. L.
,
Lose
E. J.
(
1992
).
Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans.
Annu. Rev. Cell Biol
8
,
365
393
Brennan
M. J.
,
Oldberg
A.
,
Hayman
E. G.
,
Ruoslahti
E.
(
1983
).
Effect of a proteoglycan produced by rat tumor cells on their adhesion to fibronectin-collagen substratum.
Cancer Res
43
,
4302
4307
Brown
T. A.
,
Bouchard
T.
,
John
T. St.
,
Wayner
E.
,
Carter
W. G.
(
1991
).
Human Keratinocytes express a new CD44 core protein (CD44E) as a heparan-sulfate intrinsic membrane proteoglycan with additional exons.
J. Cell Biol
113
,
207
222
Burridge
K.
,
Fath
K.
,
Kelly
T.
,
Nuckolls
G.
,
Turner
C.
(
1988
).
Focal adhesions: Transmembrane junctions between the extracellular Matrix and the cytoskeleton.
Annu. Rev. Cell Biol
4
,
487
525
Chiquet-Ehrismann
R.
,
Kalla
P.
,
Pearson
C. A.
,
Beck
K.
,
Chiquet
M.
(
1988
).
Tenascin interferes with fibronectin action.
Cell
53
,
383
390
Chiquet-Ehrismann
R.
,
Mackie
E. J.
,
Pearson
C. A.
,
Sakakura
T.
(
1986
).
Tenascin: an extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis.
Cell
47
,
131
139
Cohn
R. H.
,
Cassiman
J. J.
,
Bernfield
M. R.
(
1976
).
Relationsip of transformation, cell density and growth control to the cellular distribution of newly synthesized glycosaminoglycan.
J. Cell Biol
71
,
280
294
Couchman
J. R.
,
Austria
R.
,
Woods
A.
,
Hughes
R. C.
(
1988
).
Adhesion defective BHK cell mutant has cell surface heparan sulfate proteoglycan of altered properties.
J. Cell. Physiol
136
,
226
236
Fernandez
M. S.
,
Dennis
J. E.
,
Drushel
R. F.
,
Carrino
D. A.
,
Kimata
K.
,
Yamagata
M.
,
Caplan
A. I.
(
1991
).
The dynamics of compartmentalization of embryonic muscle by extracellular matrix molecules.
Dev. Biol
147
,
46
61
Gallagher
J. T.
(
1989
).
The extended family of PGs: Social residents of the pericellular zone.
Curr. Opin. Cell Biol
1
,
1201
1218
Grinnell
F.
(
1980
).
Visualization of cell substratum adhesion plaques by antibody exclusion.
Cell Biol. Int. Rep
4
,
1031
1036
Hasegawa
T.
,
Hasegawa
E.
,
Chen
W.-T.
,
Yamada
K. M.
) (
1985
).
Characterization of a membrane glycoprotein complex implicated in cell adhesion to fibronectin.
J. Cell. Biochem
28
,
307
318
Hayman
E. G.
,
Oldberg
A.
,
Martin
G. R.
,
Ruoslahti
E.
(
1982
).
Codistribution of heparan sulfate proteoglycan, laminin, and fibronectin in the extracellular matrix of normal rat kidney cells and their coordinate absence in transformed cells.
J. Cell Biol
94
,
28
35
Heremans
A.
,
De Cock
B.
,
Cassiman
J.-J.
,
van den Berghe
H.
,
David
G.
(
1990
).
The core protein of the matrix-associated heparan sulfate proteoglycan binds to fibronectin.
J. Biol. Chem
265
,
8716
8724
Jalkanen
M.
,
Nguyen
H.
,
Rapraeger
A.
,
Kurn
N.
,
Bernfield
M.
(
1985
).
Heparan sulfate proteoglycans from mammary epithelial cells: localization on cell surface with a monoclonal antibody.
J. Cell Biol
101
,
976
984
Kato
M.
,
Koike
Y.
,
Suzuki
S.
,
Kimata
K.
(
1988
).
Basement membrane proteoglycan in various tissues: characterization using monoclonal antibodies to the Engelbreth-Holm-Swarm mouse tumor low density heparan sulfate proteoglycan.
J. Cell Biol
106
,
2203
2210
Kimata
K.
,
Oike
Y.
,
Tani
K.
,
Sinomura
T.
,
Yamagata
M.
,
Uritani
M.
,
Suzuki
S.
(
1986
).
A large chondroitin sulfate proteoglycan (PG-M) synthesized before chondrogenesis in the limb bud of chick embryo.
J. Biol. Chem
261
,
13517
13525
Kjellen
L.
,
Lindahl
U.
(
1991
).
Proteoglycans: structures and interations.
Annu. Rev. Biochem
60
,
443
475
Knox
P.
,
Wells
P.
(
1979
).
Cell adhesion and proteoglycans. I. The effect of exogenous proteoglycans on the attachment of chick embryo fibroblasts to tissue culture plastic and collagen.
J. Cell Sci
40
,
77
88
Knudson
C. B.
,
Toole
B. P.
(
1985
).
Fluorescent morphological probe for hyaluronate.
J. Cell Biol
100
,
1753
1758
Koda
J. E.
,
Bernfield
M.
(
1984
).
Heparan sulfate proteoglycan from mouse mammary epithelial cells- basel extracellular proteoglycan binds specifically to native type I collagen fibrils.
J. Biol. Chem
259
,
11763
11770
Koda
J. E.
,
Rapraeger
A.
,
Bernfield
M.
(
1985
).
Heparan sulfate proteoglycans from mouse mammary epithelial cells. Cell surface proteoglycan as a receptor for interstitial collagens.
J. Biol. Chem
260
,
8157
8162
LeBaron
R. G.
,
Esko
J. D.
,
Woods
A.
,
Johansson
S.
,
Hook
M.
(
1988
).
Adhesion of glycosaminoglycan-deficient chinese hamster ovary cell mutants to fibronectin substrata.
J. Cell Biol
106
,
945
952
Lewandowska
K.
,
Choi
H. U.
,
Rosenberg
L. C.
,
Zardi
L.
,
Culp
L. A.
(
1987
).
Fibronectin-mediated adhesion of fibroblasts: inhibition by dermatan sulfate proteoglycan and evidence for a cryptic glycosaminoglycan-binding domain.
J. Cell Biol
105
,
1443
14547
Lightner
V. A.
,
Erickson
H. P.
(
1990
).
Binding to hexabrachion (tenascin) to the extracellular matrix and substratum and its effect on cell adhesion.
J. Cell Sci
95
,
263
277
Lories
V.
,
Cassiman
J. J.
,
van den Berghe
H.
,
David
G.
(
1992
).
Differential expression of cell surface heparan sulfate proteoglycans in human mammary epithelial cells and lung fibroblasts.
J. Biol. Chem
267
,
1116
1122
Murphy-Ullrich
J. E.
,
Lightner
V. A.
,
Aukhil
I.
,
Yan
Y. Z.
,
Erickson
H. P.
,
Hook
M.
(
1991
).
Focal adhesion integrity is down regulated by the alternatively spliced domain of human tenascin.
J. Cell Biol
115
,
1127
1136
Ninomiya
Y.
,
Nagai
Y.
(
1979
).
Modulation of glycosaminoglycan synthesis during cell growth as observed in as embryonic chick tendon cell culture.
J. Biochem. (Tokyo)
86
,
111
119
Noonan
D. M.
,
Fulle
A.
,
Valente
P.
,
Cai
S.
,
Horigan
E.
,
Sasaki
M.
,
Yamada
Y.
,
Hassell
J. R.
(
1991
).
The complete sequence of perlecan, a basemant membrane heparan sulfate proteoglycan, reveals extensive similarity with laminin a chain, low density lipoprotein-receptor, and the neural cell adhesion molecule.
J. Biol. Chem
266
,
22939
22947
Rapraeger
A.
,
Jalkanen
M.
,
Bernfield
M.
(
1986
).
Cell surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells.
J. Cell Biol
103
,
2683
2696
Rich
A. M.
,
Pearlstein
E.
,
Weissmann
G.
,
Hoffstein
S. T.
(
1981
).
Cartilage proteoglycans inhibit fibronectin-mediated adheshion.
Nature
293
,
224
226
Ruoslahti
E.
(
1989
).
Proteoglycans in cell regulation.
J. Biol. Chem
264
,
13369
13372
Saga
S.
,
Hamaguchi
M.
,
Hoshino
M.
,
Kojima
K.
(
1985
).
Expression of meta-vinculin associated with differentiation of chicken embryonal muscle cells.
Exp. Cell Res
156
,
45
56
Sanders
S.
,
Bernfield
M.
(
1988
).
Cell surface proteoglycan binds mouse mammary epithelial cells to fibronectin and behaves as a recepter for interstitial matrix.
J. Cell Biol
106
,
423
430
Singer
I. I.
,
Scott
S.
,
Kawka
D. W.
,
Hassell
J. R.
(
1987
).
Extracellular matrix fibers containing fibronectin and basement membrane heparan sulfate proteoglycan coalign with focal contacts and microfilament bundles in stationary fibroblasts.
Exp. Cell Res
173
,
558
571
Sobue
M.
,
Nakashima
N.
,
Fukatsu
T.
,
Nagasaka
T.
,
Fukuta
S.
,
Ohiwa
N.
,
Nara
Y.
,
Ogura
T.
,
Katoh
T.
,
Takeuchi
J.
(
1989
).
Production and immunohistochemical characterization of a monoclonal antibody raised to proteoglycan purified from a human yolk sac tumor.
Histochem. J
21
,
455
460
Winnemoller
M.
,
Schmidt
G.
,
Kresse
H.
(
1991
).
Influence of decorin on fibroblast adhesion to fibronectin.
Eur. J. Cell Biol
54
,
10
17
Woods
A.
,
Couchman
J. R.
(
1988
).
Focal adhesions and cell-matrix interactions.
Coll. Rel. Res
8
,
155
182
Yada
T.
,
Arai
M.
,
Suzuki
S.
,
Kimata
K.
(
1992
).
Occurrence of collagen and proeteoglycan forms of type IX collagen in chick embryo cartilage. Production and characterization of a collagen form-specific antibody.
J. Biol. Chem
267
,
9391
9397
Yamagata
M.
,
Kimata
K.
,
Oike
Y.
,
Tani
K.
,
Maeda
,
Yoshida
K.
,
Shimomura
Y.
,
Yoneda
M.
,
Suzuki
S.
(
1987
).
A monoclonal antibody that specifically recognizing a glucuronic acid 2-sulfate containing determinant in intact chondroitin sulfate.
J. Biol. Chem
262
,
4146
4152
Yamagata
M.
,
Shinomura
T.
,
Kimata
K.
(
1993
).
Tissue variation of two large chondroitin sulfate proteoglycans (PG-M/versican and PG-H/aggrecan) in chick embryos.
Anat. Embryol
187
,
433
444
Yamagata
M.
,
Suzuki
S.
,
Akiyama
S. K.
,
Yamada
K. M.
,
Kimata
K.
(
1989
).
Regulation of cell-substratum adhesion by proteoglycans immobilized on extracellular substrata.
J. Biol. Chem
264
,
8012
8018
Yamagata
M.
,
Yamada
K. M.
,
Yoneda
M.
,
Suzuki
S.
,
Kimata
K.
(
1986
).
Chondroitin sulfate proteoglycan (PG-M-like proteoglycan) is involved in the binding o hyaluronic acid to cellular fibronectin.
J. Biol. Chem
261
,
13526
13535
Yoneda
M.
,
Shimizu
S.
,
Nishida
Y.
,
Yamagata
M.
,
Suzuki
S.
,
Kimata
K.
(
1988
).
Hyaluronic acid-dependent change in the extracellular matrix of mouse dermal fibroblasts that is conductive to cell proliferation.
J. Cell Sci
90
,
275
286
Zimmermann
D. R.
,
Ruoslahti
E.
(
1989
).
Multiple domains of the large fibroblast proteoglycan, versican.
EMBO J
8
,
2975
2981
This content is only available via PDF.