Quantitation of carbon flow through the hepatic folate-dependent one-carbon pool in rats

The quantitation of carbon flow through the folate-dependent one-carbon pool in regard to the synthesis of methionine from the amino acid precursor, serine, was determined in rat liver. Utilizing duodenal cannulated rats and in vivo tracer kinetic techniques where [3-14C]serine was continuously infused at a rate of 3.32 microCi/h, a steady-state (plateau) specific radioactivity was achieved within 200 min from the onset of the infusion period. This resulted in an irreversible loss rate of 431 +/- 12 mumol/h for hepatic serine. In conjunction with the specific radioactivity measurements of hepatic methionine, the percentage of the total entry into the hepatic methionine methyl carbon pool that came from serine (i.e., transfer quotient) was calculated to be 51.7 +/- 5.2%. Similar experiments utilizing [methyl-3H]methionine as the infusate resulted in a value of 112 +/- 6 mumol/h for the irreversible loss rate of hepatic methionine. Using the irreversible loss rate of methionine and the transfer quotient to methionine from serine, the flux of the beta-carbon of serine to remethylate homocysteine and generate methionine was calculated to be 57.9 mumol/h. These results not only present a methodology for the determination of folate-dependent carbon flow in vivo, but also demonstrate the high degree to which the homocysteine moiety of methionine is conserved in vivo to meet the methylation requirements in the rat.