Glomerular epithelial cells secrete a glomerular basement membrane-degrading metalloproteinase.
Cultured rat glomerular epithelial cells (GEC) were examined for their ability to release extracellular matrix-degrading proteinases with [3H]gelatin as substrate. GEC-conditioned media, under serum-free conditions, contained modest amounts of gelatinase activity (1 to 10 U/mg of protein); the activity was maximal at neutral pH, was inhibited by zinc chelators, was not inhibited by tissue inhibitor of metalloproteinase-2, and could not be further activated by trypsin or organomercurials. Gelatin substrate sodium dodecyl sulfate-polyacrylamide gels of GEC-conditioned medium revealed several zones of lysis, with molecular sizes of 150 kd (major band), and 220, 86 to 93, and 52 to 54 kd (minor bands). Northern blot analysis demonstrated that the GEC metalloproteinase(s) were distinct from the 68- to 72-kd type IV collagenase/gelatinase present in mesangial cells or the 92-kd type IV collagenase present in neutrophils. The GEC gelatinolytic activity also degraded insoluble type IV collagen in glomerular basement membrane in a dose-dependent manner. The major metalloproteinase activity responsible for the type IV collagen degradation has a molecular size of 150 kd with a type IV collagen substrate gel. Thus, GEC produce several neutral metalloproteinases, which, by virtue of their substrate specificity, may play an important role in glomerular basement membrane remodeling and in glomerular diseases characterized by alterations in basement membrane permeability.
Johnson
R
R
Department of medicine, University of Washington Medical Center, Seattle.
Yamabe
H
H
Chen
Y P
YP
Campbell
C
C
Gordon
K
K
Baker
P
P
Lovett
D
D
Couser
W G
WG
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000201:enzymology* / Q000378:metabolism; D008666:Metalloendopeptidases / Q000145:classification / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats
D016326:Extracellular Matrix Proteins; D012333:RNA, Messenger; D003094:Collagen; D008666:Metalloendopeptidases