Localization of the cleavage sites on fibronectin following digestion by urokinase.
Urokinase (u-PA) proteolytically cleaves both human plasma (pFn) and cellular (cFn) dimeric fibronectin (M(r) 440,000) into four major polypeptides of approximately M(r) 210,000, 200,000, 25,000, and 6,000. Amino acid sequence analysis of the polypeptide fragments indicated that the enzymatic cleavage of Fn occurs at two sites: 1) between an arginine/alanine peptide bond located C-terminal to residue 259; this cleavage liberates the N-terminal M(r) 25,000 fragment and the M(r) 210,000 and M(r) 200,000 polypeptides derived from the A and B chains of Fn, respectively; and 2) between an arginine/threonine peptide bond located C-terminal to residue 2,299, thereby yielding an M(r) 6,000 dimeric fragment containing the C-terminal interchain disulfide bonds. Predigestion of Fn with u-PA increased the molecule's vulnerability to further attack by the enzymes plasmin and cathepsin D. These data provide further biochemical evidence for the proteolytic cleavage of fibronectin by plasminogen activators and substantiate that u-PA digestion of Fn may be an initial event in the local degradation of the extracellular matrix by malignant cells, possessing elevated levels of these enzymes.
Gold
L I
LI
Department of Pathology, New York University Medical Center, New York 10016.
Rostagno
A
A
Frangione
B
B
Passalaris
T
T
D000595:Amino Acid Sequence; D001665:Binding Sites; D015153:Blotting, Western; D002402:Cathepsin D / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000378:metabolism; D005341:Fibrinolysin / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D004220:Disulfides; D005353:Fibronectins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; D002402:Cathepsin D