Characterization of thrombospondin as a substrate for factor XIII transglutaminase

Thrombin activation of platelets induces the release of a high molecular weight glycoprotein, thrombospondin. On treatment with factor XIII transglutaminase and [3H]putrescine, thrombospondin undergoes specific incorporation of this labeled amine, with 2-3 mol of putrescine being incorporated per mo...

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Bibliographic Details
Published in:The Journal of biological chemistry 1987-02, Vol.262 (4), p.1772-1778
Main Authors: Lynch, G W, Slayter, H S, Miller, B E, McDonagh, J
Format: Article
Language:English
Subjects:
Autoradiography
Biological and medical sciences
Blood coagulation. Blood cells
Coagulation factors
Edetic Acid - pharmacology
Electrophoresis, Polyacrylamide Gel
Factor XIII - metabolism
Fibrin - analysis
Fibrinolysin - metabolism
Fundamental and applied biological sciences. Psychology
Glycoproteins - metabolism
Humans
Microscopy, Electron
Molecular and cellular biology
Molecular Weight
Putrescine - metabolism
Thrombospondins
Transglutaminases - metabolism
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Summary:Thrombin activation of platelets induces the release of a high molecular weight glycoprotein, thrombospondin. On treatment with factor XIII transglutaminase and [3H]putrescine, thrombospondin undergoes specific incorporation of this labeled amine, with 2-3 mol of putrescine being incorporated per mol of thrombospondin. Analysis of plasmin digests of [3H]putrescine-thrombospondin showed that the Mr 53,000-core peptide contains the glutamine site for amine incorporation. In the absence of amine substrate, thrombospondin was found to provide both donor (glutamine) and acceptor (lysine) sites for intermolecular cross-links by factors XIIIa, and high molecular weight protein complexes were formed. Homopolymers of thrombospondin were also observed by electron microscopy. Thrombin-cleaved thrombospondin has more cross-linking sites accessible for [3H]putrescine incorporation or for cross-linkage to itself than does the uncleaved native protein. Examination of thrombospondin cross-linkage in the presence of other protein substrates (fibronectin, collagen, fibrinogen, and von Willebrand factor) for factor XIIIa, resulted in reduced thrombospondin polymer formation. Electron microscopy and autoradiography of fibrin clots formed in the presence of 125I-thrombospondin showed an association of thrombospondin with fibrin fibrils. However, confirmation that this association involves covalent epsilon-(gamma-glutamyl)lysyl cross-links between thrombospondin and fibrin was not obtained.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)75705-6