A membrane-bound metallo-endopeptidase from rat kidney: its immunological characterization.
The structure and location of a membrane-bound metallo-endopeptidase, previously purified from rat kidney [Yamaguchi et al. (1991) Eur. J. Biochem. 200, 563-571], were examined by immunochemical and immunohistochemical methods with a rabbit polyclonal antibody against the purified enzyme. On treatment with endoglycosidase F, the subunit of the purified enzyme (molecular mass = 88 kDa) was converted to a smaller form (78.5 kDa), indicating that the enzyme contained at least 11% N-linked carbohydrate. Treatment of kidney membranes with papain resulted in release of the enzyme, as shown by Western blotting analysis of the solubilized fraction. Immunoassays of rat tissues showed that only the kidney, and small and large intestine expressed significant amounts of the antigen. Moreover, immunohistochemical studies showed that the antigen was confined to the luminal surfaces of the proximal renal tubules and the intestinal villi. Thus, like another kidney membrane metallo-endopeptidase, meprin [Kounnas et al. (1991) J. Biol. Chem. 266, 17350-17357], the purified enzyme is shown to be a glycoprotein that is probably anchored in the plasma membrane, and located in the luminal surface of microvillar membranes of the kidney and intestine. These results indicate that our enzyme and meprin have clear structural and topological similarities.
Yamaguchi
T
T
Department of Medicine, Kobe University School of Medicine.
Kido
H
H
Kitazawa
R
R
Kitazawa
S
S
Fukase
M
M
Katunuma
N
N
D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D002462:Cell Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D007422:Intestines / Q000201:enzymology; D007668:Kidney / Q000201:enzymology*; D007687:Kidney Tubules, Proximal / Q000201:enzymology; D008297:Male; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D008666:Metalloendopeptidases / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D008871:Microvilli / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar; D008625:Tiopronin / Q000032:analysis
D000906:Antibodies; D008625:Tiopronin; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008666:Metalloendopeptidases