Table 2.
Heme precursors and metabolites | Labeling with [14C]glycine | Labeling with [3H]ALA | |||
---|---|---|---|---|---|
14C-labeled products | Total amount | Newly synthesized amount from glycine2_a | 3H-labeled products | Total amount | |
(pmol/mg protein per 6 hr) | (pmol/mg protein per 6 hr) | ||||
Glycine | 1,200 | ||||
ALA | 100 | 3400 | 36 | ||
Heme | 5.3 | 1500b | 180 | 1.2 | 1,500b |
CO | 2.9 | 16002_c | 95 | ||
Bilirubin | 0.23 | 5702_c |
Primary cultures of olfactory receptor neurons were incubated with [14C]glycine or [3H]ALA for 6 hr. Then cellular uptakes of these precursors and biosynthesis of [14C]ALA, [14C]heme, 14CO, [3H]heme, and [3H]bilirubin were measured in these cell cultures.
Taking into account the endogenous free glycine level in neuron cells (20 nmol/mg protein), total amounts of newly synthesized products from glycine were calculated as described below. Average concentration of cellular [14C]glycine was 600 pmol/mg protein. Newly synthesized amount from glycine = 14C-labeled amount × (total glycine)/(average [14C]glycine) bThe cellular heme concentration was determined spectrophotometrically and expressed as pmol/mg protein.
The total amount of production was calculated as described below. Total amount of production=14C (or3H)-labeled production×(total heme)/[average [14C]heme (or [3H]heme)]