The determination of myeloperoxidase activity in liver.
Myeloperoxidase (MPO) is an enzyme found in granulocytes of neutrophils, but not in mammalian tissues. Previous studies have directly correlated MPO activity with neutrophil accumulation in tissues. This study presents a method for determining MPO activity in liver. Neutrophil accumulation in rat liver was provoked by creating partial ischemia followed by reperfusion. Liver homogenates prepared by a standard procedure showed no MPO activity. The homogenate was applied to Sephadex G100 and DEAE Sepharose CL6B columns which separated MPO activity from inhibitory activity. The inhibitor was identified as catalase based upon its elution from the columns and removal with 3-amino- 1,2,4-triazole (AT), a catalase inhibitor. Based upon these findings, it was determined that full MPO activity can be assayed in unfractionated liver homogenates by first inactivating catalase with AT.
Duval
D L
DL
Department of Surgery, University of Nevada School of Medicine, Reno 89557-0046.
Howard
D
D
McCalden
T A
TA
Billings
R E
RE
D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D002374:Catalase / Q000037:antagonists & inhibitors; D002850:Chromatography, Gel; D007511:Ischemia / Q000378:metabolism; D008099:Liver / Q000098:blood supply / Q000201:enzymology*; D008297:Male; D008722:Methods; D009504:Neutrophils; D009195:Peroxidase / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D002374:Catalase; D009195:Peroxidase; D000640:Amitrole