Incorporation and degradation by kidney lysosomes of cystatin alpha injected intravenously.
Cystatin alpha was purified from Escherichia coli transfected with a recombinant cystatin alpha gene and injected into the tail vein of rats. Its fate was then followed using a sensitive enzyme immunoassay for cystatin alpha. Results showed that it was rapidly removed from the blood and taken up by the kidney. Percoll density-gradient analysis showed that it was rapidly incorporated into lysosomes in the kidney. Its level in the kidney was maximal 30 min after its injection then rapidly decreased. The activity of cathepsin H in the kidney lysosomal fraction was markedly decreased 30 min after injection of cystatin alpha but recovered rapidly. Anti-(cystatin alpha) antibody precipitated cathepsin H and anti-(cathepsin H) antibody precipitated cystatin alpha in an extract of the lysosome-rich fraction of the kidney 30 min after injection of cystatin alpha. These results indicate that cystatin alpha was taken up by kidney lysosomes, formed a complex with cathepsin H and initially decreased the activity of cathepsin H, but that later the level of cystatin alpha in the kidney rapidly decreased.
Ohshita
T
T
Division of Enzyme Chemistry, University of Tokushima, Japan.
Ike
Y
Y
Katunuma
N
N
D000818:Animals; D001192:Arylsulfatases / Q000378:metabolism; D056655:Cathepsin H; D002403:Cathepsins / Q000378:metabolism; D015891:Cystatins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D007124:Immunoenzyme Techniques; D007275:Injections, Intravenous; D007668:Kidney / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
C070572:Csta protein, rat; D015891:Cystatins; D011994:Recombinant Proteins; D001192:Arylsulfatases; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056655:Cathepsin H; C534128:Ctsh protein, rat