Highly sulfated glycosaminoglycans augment the cross-linking of vitronectin by guinea pig liver transglutaminase. Functional studies of the cross-linked vitronectin multimers.
Vitronectin (VN) is an adhesive glycoprotein with roles in the complement, coagulation, and immune systems. Many of the functions of VN are mediated by a glycosaminoglycan binding site, near its carboxyl-terminal end. In this paper, we show that the highly sulfated glycosaminoglycans (GAGs), dextran sulfate, pentosan polysulfate, and fucoidan effectively augment [14C]putrescine incorporation into VN and cross-linking of VN into high molecular multimers by guinea pig liver transglutaminase (TG). Other GAGs including heparin, low molecular weight heparin, dermatan sulfate, keratan sulfate, and the nonsulfated dextrans were ineffective in accelerating these reactions. Dextran sulfate of average molecular mass 500 kDa was more effective than dextran sulfate of average molecular mass 5 kDa, supporting a template mechanism of action of the GAGs, in which VN molecules align on the GAG in a conformation suitable for cross-linking. The VN multimers catalyzed by TG retained functional activity in binding [3H]heparin, platelets, and plasminogen activator inhibitor type-1 (PAI-1). [3H]Heparin bound selectively to the 65-kDa monomeric band of VN and to the multimers derived from this band. PAI-1, however, bound equally to both the 75- and 65-kDa monomeric forms of VN, suggesting that the PAI-1 binding site on VN is distinct from the GAG binding site. The interaction of GAGs with the TG-catalyzed cross-linking of VN may facilitate studies of VN structure-function relationships.
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710.
D C
DC
Sane
T L
TL
Moser
C J
CJ
Parker
D
D
Seiffert
D J
DJ
Loskutoff
C S
CS
Greenberg
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism*; D003432:Cross-Linking Reagents; D016264:Dextran Sulfate; D003911:Dextrans / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000494:pharmacology*; D006168:Guinea Pigs; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D011485:Protein Binding; D011700:Putrescine / Q000494:pharmacology; D013329:Structure-Activity Relationship; D011503:Transglutaminases / Q000378:metabolism*; D019096:Vitronectin
D001798:Blood Proteins; D003432:Cross-Linking Reagents; D003911:Dextrans; D006023:Glycoproteins; D006025:Glycosaminoglycans; D019096:Vitronectin; D006493:Heparin; D016264:Dextran Sulfate; D011503:Transglutaminases; D004229:Dithiothreitol; D011700:Putrescine