Potential proteolytic activity of fibronectin: fibronectin laminase and its substrate specificity.
The purified 190-kDa fibronectin fragment produced by cathepsin D can be spontaneously activated in the presence of CaCl2. This activation generates new proteolytic activities and also results in the formation of several subfragments. One of them exhibits the activity of FN-gelatinase that preferentially splits type I denatured collagen and fibronectin (see preceding paper). In this work we describe the purification and characterization of another fragment (25 kDa), issued from the same autodigest. This fragment may be activated to yield another proteinase, that splits preferentially laminin and denatured collagen type I. This enzyme will be referred as FN-laminase. Purified FN-laminase specifically reacted with antibodies against fibronectin. The specificity of bond cleavage by FN-laminase was studied with various synthetic peptides analogous to collagen repeats. FN-laminase cleaves the Ala-Gly bond in the sequence GPAGPR; the arginine residue in position P3' is important for this cleavage. The enzyme is inhibited by pepstatin A and phenylmethanesulfonyl fluoride, like retroviral aspartic proteinases. It is also inhibited by EDTA. No inhibition was obtained with 1,10-phenanthroline or 4-chloromercuribenzoate, inhibitors of Zn-metalloproteinases or cysteine proteinases, respectively.
Emod
I
I
Unité de Chimie des Protéines, Institut Pasteur, Paris.
Lafaye
P
P
Planchenault
T
T
Lambert Vidmar
S
S
Imhoff
J M
JM
Keil-Dlouha
V
V
D000595:Amino Acid Sequence; D003094:Collagen / Q000378:metabolism; D004789:Enzyme Activation; D005353:Fibronectins / Q000032:analysis*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007797:Laminin / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis*; D011480:Protease Inhibitors; D012190:Retroviridae / Q000201:enzymology; D013379:Substrate Specificity
D005353:Fibronectins; D007074:Immunoglobulin G; D007797:Laminin; D010446:Peptide Fragments; D011480:Protease Inhibitors; D003094:Collagen