An increase in cytosolic protease activity during liver preservation. Inhibition by glutathione and glycine.
Degradative cytosolic proteolysis contributes to cell injury following ATP depletion. Although ATP depletion is a salient feature of ischemic liver storage for transplantation, information regarding cytosolic protease activity during liver storage is lacking. Thus our aim was to measure liver cytosolic protease activity following ischemic storage. A progressive increase in total cytosolic protease activity was observed over time at both 37 degrees C and 4 degrees C, but the increase was greater at 37 degrees C. Total cellular proteolysis was also temperature-dependent during anoxia (37 degrees C > 4 degrees C), demonstrating a physiologic correlation between cellular proteolysis and measurements of cytosolic protease activity. The stimulation of total cytosolic protease activity was due to an increase in metallo- and aspartate protease activity. Of particular interest, glutathione (GSH) inhibited both metalloprotease and aspartate protease activity from cytosol of stored livers. Glycine, the carboxyl-terminal amino acid of GSH, also inhibited both metalloprotease and aspartate protease activity. In addition to being an antioxidant, GSH may exert its protective effects during organ preservation by inhibiting cytosolic proteases--perhaps via its glycine moiety. These experiments support the hypothesis that degradative proteolysis contributes to liver injury during organ preservation.
Ferguson
D M
DM
Section of Transplantation Surgery, Mayo Clinic, Rochester, Minnesota 55905.
Gores
G J
GJ
Bronk
S F
SF
Krom
R A
RA
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology*; D010450:Endopeptidases / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology*; D005998:Glycine / Q000494:pharmacology*; D006026:Glycoside Hydrolases / Q000378:metabolism; D000860:Hypoxia / Q000378:metabolism; D007511:Ischemia / Q000378:metabolism; D008099:Liver / Q000098:blood supply / Q000166:cytology; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000494:pharmacology; D009926:Organ Preservation; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats; D011917:Rats, Inbred Lew; D013696:Temperature
D011480:Protease Inhibitors; D006026:Glycoside Hydrolases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D005978:Glutathione; D005998:Glycine