title: Human plasma fibronectin as a substrate for human urokinase.
abstract: An early event in malignant transformation is the increased expression of proteases, such as plasminogen activator, which can degrade surrounding extracellular matrices, thereby conferring an advantage for tumour cell invasion and metastasis. The present studies provide evidence that plasma fibronectin (Fn), which is a component of the extracellular matrix, is a direct substrate for the plasminogen activator urokinase (UK). Human plasma Fn was incubated with human UK under plasminogen-free conditions. Fn cleavage was both time- and dose-dependent and was evident within 30 min. The proteolytic digestion was limited and complete within 12 h at an enzyme/substrate ratio of 1:20. Analysis of the final proteolytic digestion products demonstrated the disappearance of the native dimeric 440 kDa structure of Fn with the concomitant appearance of three proteolytic fragments of 210, 200 and 25 kDa. Since two large fragments of similar size to the 220 kDa monomeric chains of Fn were obtained following proteolysis, it is proposed that UK cleaves Fn at two sites, one towards the N-terminal and one close to the C-terminal, but N-terminal to its interchain disulphide bonds. These studies suggest that the local proteolytic digestion and release of Fn from the extracellular matrix by tumour cells possessing high levels of UK may involve the direct proteolytic breakdown of Fn by UK.
authors.affiliation: NYU Medical Center, Department of Pathology, New York 10016.
authors.forename: L I
authors.initials: LI
authors.lastname: Gold
authors.forename: R
authors.initials: R
authors.lastname: Schwimmer
authors.forename: J P
authors.initials: JP
authors.lastname: Quigley
mesh_terms: D005341:Fibrinolysin; D005353:Fibronectins / Q000097:blood*; D006801:Humans; D015394:Molecular Structure; D013997:Time Factors; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator