Thrombospondin interaction with plasminogen. Evidence for binding to a specific region of the kringle structure of plasminogen.
Platelet thrombospondin interacts with plasminogen in a specific and saturable manner. Thrombospondin was found to specifically bind to plasminogen and the nonenzyme chain of plasmin. Preincubation of 125I-labeled thrombospondin with 30 mmol/L lysine was without effect in the binding of thrombospondin to immobilized plasminogen; preincubation of 125I-labeled plasminogen with 30 mmol/L lysine, on the other hand, significantly reduced the binding of plasminogen to immobilized thrombospondin, suggesting that the interaction of thrombospondin with plasminogen is not the direct result of the lysine binding sites of plasminogen. Arginine and benzamidine, ligands known to specifically bind to the kringle 5 domain of plasminogen, blocked the binding of thrombospondin to plasminogen. Limited elastase proteolysis of plasminogen and plasmin resulted in the generation of two distinct thrombospondin binding domains, one of which was retained on lysine-agarose. The isolation and amino-terminal analysis of these domains following elastase proteolysis of plasminogen identified them, respectively, as a domain containing kringle structures 4 and 5 and plasmin and the other domain consisting of kringle 5-plasmin. A 16-residue synthetic peptide, which represents the amino acids linking kringle 4 to kringle 5 (residues 435-450 of native plasminogen), was without effect in either binding to thrombospondin or blocking the binding of thrombospondin to plasminogen. Plasminogen, therefore, possesses a single thrombospondin interactive site that is independent of, but influenced by, the lysine binding site containing kringle structures and most likely is located within the kringle 5 domain.
DePoli
P
P
Department of Physiology, Wayne State University, Detroit, MI 48201.
Bacon-Baguley
T
T
Kendra-Franczak
S
S
Cederholm
M T
MT
Walz
D A
DA
D000818:Animals; D002417:Cattle; D005341:Fibrinolysin / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D010958:Plasminogen / Q000378:metabolism*; D019699:Thrombospondins
D006023:Glycoproteins; D019699:Thrombospondins; D010958:Plasminogen; D005341:Fibrinolysin