Identification of latent procathepsin H in microsomal lumen: characterization of proteolytic processing and enzyme activation.
Procathepsin H in kidney and liver microsomal lumen was identified to have a molecular mass of 41 kDa by immunoblot analysis. The proenzyme was then concentrated by applying the microsomal contents to a concanavalin A-Sepharose column. When the concanavalin A-adsorbed fraction was incubated at pH 4.0 at 20 degrees C, the activity measured with synthetic substrate increased 3.5 times over that of the control after 24 h incubation. Immunoblot analysis showed that acidic treatment caused the disappearance of procathepsin H. Thus the proenzyme might be processed to the mature enzyme under acidic conditions. The marked increase of enzymatic activity and the conversion of proenzyme were completely blocked with pepstatin which is a potent inhibitor of aspartic proteases. These results suggested that a protease for processing procathepsin H might be cathepsin D, a major lysosomal aspartic protease. Therefore, procathepsin H seems to be synthesized first in the enzymatically inactive form in endoplasmic reticulum and successively converted into the active form in lysosomes during biosynthesis.
Nishimura
Y
Y
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Kato
K
K
D000818:Animals; D002402:Cathepsin D / Q000378:metabolism; D056655:Cathepsin H; D002403:Cathepsins / Q000032:analysis / Q000378:metabolism*; D002846:Chromatography, Affinity; D003546:Cysteine Endopeptidases; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000032:analysis / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000648:ultrastructure*; D008297:Male; D008861:Microsomes / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010436:Pepstatins / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D004792:Enzyme Precursors; D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D002403:Cathepsins; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D056655:Cathepsin H; C534128:Ctsh protein, rat; D002402:Cathepsin D; C031375:pepstatin