Degradation of basement membranes by human matrix metalloproteinase 3 (stromelysin).
Connective tissue cells synthesize and secrete a group of matrix metalloproteinases (MMPs), all of which are capable of degrading the extracellular-matrix components. One of them, MMP-3 (stromelysin) has been shown to degrade purified basement-membrane components, collagen IV and laminin [Okada, Y., Nagase, H. & Harris, E. D., Jr. (1986) J. Biol. Chem. 261, 14245-14255]. Here we report that MMP-3 degrades collagen IV and laminin in intact basement membranes from bovine glomeruli (GBM) and bovine anterior-lens capsules (LBM). Degradation products were analysed by SDS/polyacrylamide-gel electrophoresis to determine the number and sizes of polypeptide fragments. Immunoblotting techniques were used to identify the origins of the fragments, i.e. collagen IV or laminin. The fragments of collagen IV were further mapped using specific antibodies that recognize the N-terminal (7 S) domain, the C-terminal (NC-1) domain, or the major triple-helical region between the terminal domains. Degradation of collagen IV was extensive; many fragments were found, from both GBM and LBM, in the Mr range 25,000-380,000. A large fragment of laminin (Mr greater than 380,000) was found in the GBM digests without reduction, but it dissociated into 220,000-Mr chains upon reduction. The results suggest that MMP-3 plays an important role in the catabolism of basement membranes.
Bejarano
P A
PA
Department of Biochemistry, University of Kansas Medical Center, Kansas City 66103.
Noelken
M E
ME
Suzuki
K
K
Hudson
B G
BG
Nagase
H
H
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D015153:Blotting, Western; D002417:Cattle; D003094:Collagen / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007678:Kidney Glomerulus / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism; D007908:Lens, Crystalline / Q000378:metabolism*; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000378:metabolism*
D007797:Laminin; D003094:Collagen; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3