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J. Biol. Chem., Vol. 261, Issue 15, 6872-6877, May, 1986
Comparison of the specificities of laminin, thrombospondin, and von Willebrand factor for binding to sulfated glycolipids
DD Roberts, CN Rao, LA Liotta, HR Gralnick and V Ginsburg
The adhesive glycoproteins laminin, thrombospondin, and von Willebrand
factor bind specifically and with high affinity to sulfated glycolipids.
These three glycoproteins differ, however, in their sensitivity to
inhibition of binding by sulfated monosaccharides and polysaccharides.
Heparin strongly inhibits binding of thrombospondin but only weakly
inhibits binding of laminin and von Willebrand factor. Fucoidan strongly
inhibits binding of both laminin and thrombospondin but not of von
Willebrand factor. Laminin shows significant specificity for inhibition by
monosaccharides, whereas thrombospondin does not. Thus, specific spacial
orientations of sulfate esters may be primary determinants of binding for
the three proteins. Laminin, thrombospondin, and von Willebrand factor also
differ in their relative binding affinities for purified sulfated
glycosphingolipids. The three proteins strongly prefer terminal-sulfated
lipids and bind only weakly to sulfated gangliotriaosyl ceramide with a
sulfate ester on the penultimate galactose. Thrombospondin binds with
highest affinity to galactosyl sulfatide but only weakly to more complex
sulfatides, whereas von Willebrand factor prefers galactosyl sulfatide but
binds with moderate affinity to various sulfated glycolipids. Laminin also
is less selective than thrombospondin but is less sensitive for detection
of low sulfatide concentrations. Galactosyl sulfatide at 1-5 pmol can be
detected by staining of lipids separated on high performance TLC with
125I-thrombospondin or 125I-von Willebrand factor. 125I-von Willebrand
factor was examined as a reagent for detecting sulfated glycolipids in
tissue extracts. Rat kidney lipids contain 5 characterized sulfated
glycolipids: galactosyl ceramide I3-sulfate, lactosyl ceramide II3-sulfate,
gangliotriaosyl ceramide II3-sulfate, and bis-sulfated gangliotriaosyl and
gangliotetraosyl ceramides. von Willebrand factor detects all of these
lipids as well as several additional minor sulfated lipids. Complex
monosulfated lipids are detected in several human tissues including kidney,
erythrocytes, and platelets by this technique.

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Copyright © 1986 by the American Society for Biochemistry and Molecular Biology.
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