Elastase inhibitory capacity of purified canine alpha-1-antitrypsin.
In a previous study, isoelectrofocusing of serum from liver-diseased and healthy dogs revealed three different types of the acute phase protein, alpha-1-antitrypsin: Pi (protease inhibitor) F, Pi I and Pi S. Moreover, accumulated alpha-1-antitrypsin was found immunohistochemically in liver sections from dogs with chronic liver disease, predominantly in association with Pi I in serum. The present study was made to further the relationship between alpha-1-antitrypsin and the pathogenesis of chronic liver disease in dogs. Aliquots of samples of purified canine Pi F, Pi I and Pi S were examined for elastase inhibitory capacity, the main function of alpha-l-antitrypsin, and for polymerization tendency, a possible cause of accumulation of alpha-1-antitrypsin in the liver. These parameters were studied after incubation of the proteins at different temperatures (4, 37 and 42 degrees C) and pH values (6.8, 7.8 and 8.5) and for different periods (< or = 24 h and 5 days). In contrast to findings with Pi Z, the human alpha-1-antitrypsin variant associated with liver disease, polymers of canine Pi F, Pi I or Pi S could not be detected under any of the conditions tested. However, Pi I was sensitive to pH, as was demonstrated by reduced elastase inhibitory capacity after incubation at pH 6.8 for < or = 24 h or 5 days, or at pH 8.5 for 5 days. However, after incubation at pH 7.8 for < or = 24 h or 5 days at 4, 37 or 42 degrees C, Pi I was completely stable. Pi F retained its elastase inhibitory capacity, even after prolonged incubation, at all pH values and temperatures tested. Due to low yield, Pi S was tested only after incubation for < or = 24 h at pH 6.8 and at 4 degrees C; under these conditions its elastase inhibitory capacity was equal to that of Pi F. Taken together, these findings indicate molecular and functional differences between Pi I and Pi F and further support a role for alpha-1-antitrypsin in the pathogenesis of canine liver disease.
Vatne
M
M
Animal Hospital of Helsingborg, Small Animal Clinic, Sweden.
Andersson
M
M
Sevelius
E
E
Jönsson
L
L
D000818:Animals; D001704:Biopolymers; D002908:Chronic Disease; D004285:Dogs / Q000378:metabolism*; D005838:Genotype; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors*; D008099:Liver / Q000201:enzymology; D008107:Liver Diseases / Q000201:enzymology; D010641:Phenotype; D013696:Temperature; D000515:alpha 1-Antitrypsin / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*
D001704:Biopolymers; D000515:alpha 1-Antitrypsin; D019272:Leukocyte Elastase