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In some tissues, amino acids (AA) stimulate translation initiation via interactions between eukaryote initiation factor (eIF) 4E-binding protein 1 (4E-BP1), eIF4E and eIF4G. Dietary AA have been shown to induce pancreatic proteases independently of cholecystokinin in rats, the mechanism of which has not yet been clarified. In the present study, we examined the mechanism in rats for protease induction by dietary AA and determined the involvement of translation initiation. Male Wistar/ST rats were fed a 20 or 60% casein or AA mixture diet for 7 d and were intravenously injected with [35S] methionine (Met) 30 min before killing on d 7 (expt. 1). In expt. 2, rats were fed a 20 or 60% AA diet for 7 d and after food deprivation and refeeding with the respective diet on d 7 were killed at 0, 1 or 3 h. We measured mRNA and [35S] Met incorporation into chymotrypsinogen, phosphorylation status of 4E-BP1 and the association of eIF4E with 4E-BP1 or eIF4G. In expt. 1, chymotrypsin activity and synthesis were higher in both of the 60% diet groups than in the 20% diet groups, but the mRNA level and 4E-BP1 status did not differ. In expt. 2, chymotrypsin activity increased in the 60% AA diet group in a time-dependent manner. The translation initiation activity via the mTOR pathway indicated an increase similar to chymotrypsin activity. There were no differences in chymotrypsin mRNA level at any point. These results indicate that dietary AA induce chymotrypsin synthesis by promoting translation, and transient activation of translation initiation via mTOR may be associated with this induction.

Rats were fed raw soybeans or purified soybean proteinase inhibitors by tube to see whether they were able to produce inhibitor-resistant trypsin, as previously demonstrated in humans. Their duodenal chyme contained only 20-50% of the enzymatic activities of animals fed bovine serum albumin (BSA) as test protein. However, both tryptic and chymotryptic activities had considerable resistance to low- and high-molecular-weight inhibitors of serine proteinases. In particular, the tryptic activity demonstrated a high degree of inhibitor resistance. Human alpha 1-antitrypsin and lima bean inhibitor in amounts that inhibited bovine serum albumin-induced trypsin completely caused only 2-12% inhibition of the raw soybean-induced tryptic activity. The inhibitor-resistant tryptic and chymotryptic activities after raw soybean instillation might be caused by the Bowman-Birk and Kunitz trypsin inhibitors. The physiologic significance of an inhibitor-resistant trypsin might be to assure activation of other pancreatic proenzymes. The results of the present rat experiments confirm the previous findings of inhibitor-resistant trypsin in humans.