Purification and properties of serine hydroxymethyltransferase and C1-tetrahydrofolate synthase from L1210 cells

William B. Strong, Saul J. Tendler, Richard Seither, I. David Goldman, Verne Schirch

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Serine hydroxymethyltransferase and the trifunctional enzyme C1-tetrahydrofolate synthase have been purified to near homogeneity from L1210 cells. Kinetic constants (Km and kcat) have been determined for both folate and non-folate substrates. The effect of increasing glutamate chain length on affinity and catalytic efficiency were determined for the four activities. The studies show that the structural and catalytic properties of the two L1210 enzymes are very similar to the corresponding enzymes purified from rabbit liver. Antibodies to both rabbit serine hydroxymethyltransferase and C1-tetrahydrofolate synthase cross-react with the corresponding L1210 enzymes. The intracellular concentration of active sites of serine hydroxymethyltransferase and C1-tetrahydrofolate synthase in L1210 cells are both 9 μM. The combined concentration of these two enzymes exceeds the previously reported concentration of 10 μM for total intracellular folates. A network thermodynamic computer model of one carbon metabolism (Seither, R. L., Trent, D. F., Mikulecky, D. C., Rape, T. J., and Goldman, I. D. (1989) J. Biol. Chem. 264, 17016-17023) suggests that complete inhibition of cytosolic serine hydroxymethyltransferase would neither significantly decrease the rates of biosynthesis of purines and thymidylate nor significantly alter the rate of interconversion of tetrahydrofolate cofactors to dihydrofolate with subsequent inhibtion of dihydrofolate reductase.

Original languageEnglish (US)
Pages (from-to)12149-12155
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number21
StatePublished - Jul 25 1990
Externally publishedYes

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Glycine Hydroxymethyltransferase
Purification
Enzymes
Folic Acid
Rabbits
Purines
Tetrahydrofolate Dehydrogenase
Biosynthesis
Chain length
Thermodynamics
Metabolism
Liver
Computer Simulation
Glutamic Acid
Catalytic Domain
Carbon
formyl-methenyl-methylenetetrahydrofolate synthetase
Kinetics
Antibodies
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and properties of serine hydroxymethyltransferase and C1-tetrahydrofolate synthase from L1210 cells. / Strong, William B.; Tendler, Saul J.; Seither, Richard; Goldman, I. David; Schirch, Verne.

In: Journal of Biological Chemistry, Vol. 265, No. 21, 25.07.1990, p. 12149-12155.

Research output: Contribution to journalArticle

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