Structural basis of beta 1 integrin-mediated cell adhesion to a large heparan sulfate proteoglycan from basement membranes.
In a cell attachment assay, several cell lines were found to adhere and spread on heparan sulfate proteoglycan purified from a basement membrane-producing tumor. This adhesion was clearly distinct from that on laminin. Cell adhesion to the proteoglycan was completely inhibited by three different antibodies against integrin beta 1 subunit, while inhibitory antibodies against beta 3 and alpha 2 to alpha 6 subunits were without strong effects. Removal of heparan sulfate from the proteoglycan diminished cell attachment, but addition of heparin to the cells did not affect adhesion to the proteoglycan. This suggests that both the heparan sulfate side chains and core protein structures are required for efficient cell adhesion. Studies with proteolytic fragments and synthetic RGD peptides showed that the single RGD sequence of mouse proteoglycan is not involved in cellular recognition. Characterization of fragments also allowed to localize cell adhesion and heparan sulfate attachment sites to the same 160 kDa core protein structure but not to fragments derived from the N-terminal portion of the proteoglycan.
Battaglia
C
C
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Aumailley
M
M
Mann
K
K
Mayer
U
U
Timpl
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001485:Basement Membrane / Q000737:chemistry; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000502:physiology*; D019012:Integrin beta1; D016023:Integrins / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011509:Proteoglycans / Q000502:physiology*
D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D019012:Integrin beta1; D016023:Integrins; D010446:Peptide Fragments; D011509:Proteoglycans