Serine hydroxymethyltransferase (SHMT) is the primary enzyme in the interconversion of serine and glycine. The roles of mitochondrial and cytosolic SHMT in the interconversion of serine and glycine were determined in two Chinese hamster ovary (CHO) cell lines that both contain cytosolic SHMT but either have (CHOm+) or lacK (CHOm-) mitochondrial SHMT. Mitochondrial SHMT activity was significantly reduced in CHOm- (0.24 +/- 0.11 nmol/min per mg of mitochondrial protein) compared with CHOm+ (3.21 +/- 0.66 nmol/min per mg of mitochondrial protein; P = 0.02) cells, whereas cytosolic SHMT activity was similar in CHOm- and CHOm+ cells (1.09 +/- 0.31 and 1.53 +/- 0.12 nmol/min per mg of cytosolic protein respectively; P = 0.57). In CHOm+ and CHOm- cells, the relative flux of glycine to serine measured with either [1-13C]- or [2-13C]-glycine was similar (CHOm-: 538 +/- 82 nmol/24 per mg of DNA; CHOm+: 616 +/- 88 nmol/24 h per mg of DNA; P = 0.42). In contrast, the relative flux of serine to glycine measured with [1-13C]serine was low in CHOm- cells (80 +/- 28 nmol/24 h per mg of DNA) compared with CHOm+ cells (3080 +/- 320 nmol/24 h per mg of DNA; P = 0.0001). The rate of glycine production determined by [1-(13)C]glycine dilution was lower in CHOm- (1200 +/- 200 nmol/24 h per mg of DNA) than CHOm+ (10200 +/- 1800 nmol/24 h per mg of DNA; P = 0.03) cells, whereas glycine utilization was similar in the two cell lines. Serine production was similar in the two cell lines but serine utilization was lower in CHOm- (3800 +/- 1200 mu mol/24 h per mg of DNA) than CHOm+ (6600 +/- 1000 nmol/24 h per mg of DNA; P = 0.0002) cells. Increasing the serine concentration in the medium resulted in an increase in glycine production in CHOm+ but not in CHOm- cells. Intracellular studies with [1-13C]serine confirm the findings of decreased glycine production from serine. In CHO cells there is partitioning of intracellular serine and glycine metabolism. Our data support the hypothesis that mitochondrial SHMT is the primary pathway for serine into glycine interconversion.

Using stable isotopes of serine, glycine, and glutamine, the metabolism of serine and glycine was investigated in primary hepatocytes from six mid-gestation fetal lambs (mean gestational age = 81 +/- 6 d, normal gestation = 145 d). Serine production was 6.84 +/- 1.22 mumol/24 h/mg of DNA and exceeded serine utilization (3.76 +/- 1.44 mumol/24 h/mg of DNA) with a resultant net increase in medium serine of 2.58 +/- 1.70 mumol/24 h/mg of DNA. In contrast, glycine production (6.84 +/- 1.16 mumol/24 h/mg of DNA) was less than glycine utilization (12.10 +/- 1.78 mumol/24 h/mg of DNA) with a net decline in medium glycine of -5.44 +/- 2.03 mumol/24 h/mg of DNA. Of the serine produced, 50.4 +/- 4.3% was derived from glycine via the action of serine hydroxymethyltransferase (SHMT) and the glycine cleavage enzyme complex (GCS). Increasing the medium serine concentration resulted in an increase in serine utilization and sparing of the utilization of other amino acids. Biosynthesis of glycine from serine accounts for only 18.1 +/- 5.6% of glycine production, and this percentage is not affected by changes in medium serine concentration. Using 2.5-[15N2]glutamine as the tracer, an estimated 18 +/- 7% of serine production was derived from transamination reactions. The specific activity of both cytosolic and mitochondrial SHMT was constant for the duration of the cultures. We conclude that, in mid-gestation fetal ovine hepatocytes, there is net production of serine (with glycine as the primary metabolic source of this serine biosynthesis) and net glycine utilization. These data suggest that flux through SHMT and GCS accounts for 50% of serine biosynthesis in mid-gestation fetal ovine hepatocytes. The sparing of the utilization of other amino acids by serine suggests that serine a conditionally essential amino acid for the mid-gestation fetal liver.