Evidence for intracellular partitioning of serine and glycine metabolism in Chinese hamster ovary cells

Abstract

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.

Ternary complex formation and competition quench fluorescence of ZnAF family zinc sensors

“…Histidine and 60 glycine as representative amino acids of the protein biosynthesis pool, vary according to cell cycle and metabolism. 15,16 GSH is synthesized and used up according to various cellular stresses and assaults. 17,18 These compounds also represent a variety of Zn(II) binding Roth, citric acid hydrate, 99%, from Standard, Poland, MgCl 2 hexahydrate, 99%, from Merck.…”

Integrated Temporal Regulation of the Photorespiratory Pathway. Circadian Regulation of Two Arabidopsis Genes Encoding Serine Hydroxymethyltransferase

“…Methylene THF can be converted to methyl-, methenyl-and formyl-THF, which serve as the primary sources of onecarbon (C 1 ) units of differing oxidation states for biosynthetic reactions in amino acid, purine, pyrimidine, and lipid synthesis (Cossins, 1987). Eukaryotes have both cytosolic and mitochondrial SHMT isoforms (Cossins, 1987), and evidence is accumulating for the intracellular partitioning of Ser and Gly metabolism (Narkewicz et al, 1996;Kastanos et al, 1997).…”

Serine Hydroxymethyltransferase Isoforms Are Differentially Inhibited by Leucovorin: Characterization and Comparison of Recombinant Zebrafish Serine Hydroxymethyltransferases

“…In higher organisms more than one SHMT isoform is often present: a cytosolic isoform and an organelleassociated form, usually mitochondria. Although the physiological functions of these two isozymes remain unclear, it is suggested that cytosolic SHMT (cSHMT) tends to function in the direction of serine synthesis, whereas the production of glycine and N 5 ,N 10 -CH 2 -THF is mainly catalyzed by mitochondrial SHMT (mSHMT) (Narkewicz et al, 1996). Studies suggested that the activity of cSHMT is associated with the development of MTX resistance.…”