13C enrichment of carbons 2 and 8 of purine by folate-dependent reactions after [13C]formate and [2-13C]glycine dosing in adult humans.

The 10-formyl moiety of 10-formyltetrahydrofolate is the source of carbons at the positions 8 (C(8)) and 2 (C(2)) of the purine ring, originating from formate and a few amino acids. Uric acid is the final catabolic product of purines. In adult humans, we independently measured the (13)C enrichment of the C(2) and C(8) positions of urinary uric acid after an oral dose of [(13)C]sodium formate and that of the C(2) and C(8) plus C(5) positions after [2-(13)C]glycine. A liquid chromatography-mass spectrometric method was used to measure the (13)C enrichment of uric acid in urine, which was collected for 3 to 4 days. Purine catabolism to uric acid does not alter the positions of carbons in the ring. After the formate dose, the (13)C enrichment at C(2) was greater than at C(8), and a circadian rhythm was observed in the enrichment at C(2). After the glycine dose, the C(8) plus C(5) positions were enriched, whereas no significant enrichment at C(2) was found. These (13)C enrichment patterns are not consistent with previous accepted metabolism. To our knowledge, this is the first study to investigate (13)C enrichment from formate and glycine independently into the C(2) and C(8) positions of purine in the same subjects. Possible mechanisms explaining our findings are discussed. Oral [(13)C]formate or [2-(13)C]glycine dosing and urine collection can be used to study purine biosynthesis in humans.