Compartmentation of folate metabolism in rat pancreas: nitrous oxide inactivation of methionine synthase leads to accumulation of 5-methyltetrahydrofolate in cytosol.

Folate-dependent one-carbon metabolism and methylation reactions have been implicated in the secretory function of the pancreas. Because vitamin B-12 deficiency perturbs folate metabolism, we determined the effects of nitrous oxide inactivation of methionine synthase on the compartmentation of folate metabolism in rat pancreas. Rats were exposed to an atmosphere of nitrous oxide and oxygen (80 and 20%, respectively) for 18 h; control rats breathed air. Folate coenzyme concentrations were determined by HPLC and Lactobacillus casei microbiological assay of the cytosolic and mitochondrial fractions of pancreas, which contained 62 and 46%, respectively, of the total folate. In pancreas of control rats, cytosolic folates were 5-methyltetrahydrofolate (31% of total folates), tetrahydrofolate (54%) and 5- and 10-formyltetrahydrofolate (6 and 8%, respectively). In the rats exposed to nitrous oxide, cytosolic 5-methyltetrahydrofolate concentrations were significantly greater (59% of total folates) and tetrahydrofolate concentrations were significantly lower (32%) than in controls; however, total cytosolic folate levels were unaffected by nitrous oxide exposure. In controls, mitochondrial folates were composed of 5-methyltetrahydrofolate (9% of total folates), tetrahydrofolate (60%) and 5- and 10-formyltetrahydrofolate (22 and 10%, respectively). Exposure to nitrous oxide led to significantly lower total mitochondrial folates (1.49 +/- 0.18 vs. 0.75 +/- 0.29 nmol/g, control vs. nitrous oxide, P < 0.05). This was due to a significantly lower concentration of tetrahydrofolate and 5-formyltetrahydrofolate, but not of 5-methyl- or 10-formyltetrahydrofolate. The activity of methionine synthase was 85% lower (P < 0.001) in pancreatic extracts of rats exposed to nitrous oxide than in controls. These results show that cytosolic folates accumulate in pancreas as the 5-methyl derivative at the expense of other reduced folates, as happens in liver. However, in contrast to results in liver, the mitochondrial folate concentration was lower in the pancreas of rats exposed to nitrous oxide, and this decline was limited to the 5-formyl- and tetrahydrofolate derivatives.