Identification of fibronectin type I domains as amyloid-binding modules on tissue-type plasminogen activator and three homologs

Abstract

The serine protease tissue-type plasminogen activator (tPA), a key enzyme in hemostasis, is activated by protein aggregates with amyloid-like properties. tPA is implicated in various pathologies, including amyloidoses. A major task is to further elucidate the mechanisms of amyloid pathology. We here show that the fibronectin type I domain of tPA mediates the interaction with amyloid protein aggregates. We found that in contrast to full-length tPA, a deletion-mutant of tPA, lacking the first three N-terminal domains (including the fibronectin type I domain), fails to activate in response to amyloid protein aggregates. Using recombinantly produced domains of tPA in direct binding assays, we subsequently mapped the amyloid-binding region to the fibronectin type I domain. This domain co-localized with congophylic plaques in brain sections from patients with Alzheimer's disease. Fibronectin type I domains from homologous proteases factor XII, hepatocyte growth factor activator and from the extracellular matrix protein fibronectin also bound to aggregated amyloidogenic peptides. Finally, we demonstrated that the isolated fibronectin type I domain inhibits amyloid-induced aggregation of blood platelets. The identification of the fibronectin type I domain as an amyloid-binding module provides new insights into the (patho-) physiological role of tPA and the homologous proteins which may offer new targets for intervention in amyloid pathology.

Keywords:

• tPA
• amyloid
• fibrinolysis
• fibronectin type I domain

Abbreviations
tPA	=	tissue-type plasminogen activator;
FXII	=	coagulation factor XII;
HGFA	=	hepatocyte growth factor activator;
Fn	=	fibronectin;
Aβ	=	amyloid β peptide;
ATTR	=	transthyretin;
ELISA	=	enzyme-linked immunosorbent assay;
F	=	fibronectin type I (finger) domain;
EGF	=	epidermal growth factor;
E	=	EGF-like domain;
K	=	kringle domain;
εACA	=	epsilon-amino caproic acid;
BSA	=	bovine serum albumin;
AGE	=	advanced glycation end-product;
GST	=	glutathione-S-transferase;
LAM12	=	laminin α1-chain (2097–2108) amyloid core peptide

Abbreviations
tPA	=	tissue-type plasminogen activator;
FXII	=	coagulation factor XII;
HGFA	=	hepatocyte growth factor activator;
Fn	=	fibronectin;
Aβ	=	amyloid β peptide;
ATTR	=	transthyretin;
ELISA	=	enzyme-linked immunosorbent assay;
F	=	fibronectin type I (finger) domain;
EGF	=	epidermal growth factor;
E	=	EGF-like domain;
K	=	kringle domain;
εACA	=	epsilon-amino caproic acid;
BSA	=	bovine serum albumin;
AGE	=	advanced glycation end-product;
GST	=	glutathione-S-transferase;
LAM12	=	laminin α1-chain (2097–2108) amyloid core peptide