Glucagon degradation in fraction of big plasma glucagon in rats treated with carbon tetrachloride.
When whole plasma obtained from rats given carbon tetrachloride was applied to a column of Bio Gel P-30, P-150 or P-300, a large molecular form of immunoreactive glucagon (IRG), so called "big plasma glucagon" (BPG), was eluted in the void volume. Glucagon degrading activity, measured under the same conditions as for radioimmunoassay of IRG, was eluted in parallel with IRG. Since standard assay mixture containing aprotinin was used for the measurement of IRG, this degradation of glucagon might be due to an enzyme(s) other than serine proteinases. p-Hydroxymercuribenzoate depressed glucagon degradation and IRG in the BPG fractions to 20% and 30% of the control values, respectively. Other inhibitors of thiol proteinases, such as N-ethylmaleimide and leupeptin, also decreased the values for both glucagon degradation and IRG to a similar extent. However, phenylmethylsulfonyl fluoride and pepstatin had little or no effect on these values. On filtration of plasma from carbon tetrachloride-treated rats on a column of Bio Gel A-1.5m, IRG peaks between 600k and 1,200k daltons were obtained and glucagon degrading activity was again eluted with IRG. When p-chloromercuriphenyl sulfonate and N-ethylmaleimide were added to the elution buffer and assay mixtures, the glucagon degrading activity was almost completely inhibited and the values for IRG between 600k and 1,200k daltons were decreased to less than 10% of those of the control. These results indicate that more than 90% of the IRG in the fractions of BPG obtained from carbon tetrachloride-treated rats could be accounted for entirely by the degradation of immunoreactive 125I-glucagon by plasma proteinases, probably thiol proteinases. Thus we conclude that some proteinases are still active in the radioimmunoassay system of glucagon which is widely used at present, and lead apparently higher glucagon values owing to degradation of 125I-glucagon.
D000818:Animals; D002251:Carbon Tetrachloride / Q000494:pharmacology; D005934:Glucagon / Q000378:metabolism*; D008297:Male; D011480:Protease Inhibitors / Q000494:pharmacology; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains
D011480:Protease Inhibitors; D005934:Glucagon; D002251:Carbon Tetrachloride