Publications

• Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase.

  Inglese J., Johnson D.L., Shiau A., Smith J.M., Benkovic S.J.

  Glycinamide ribonucleotide transformylase (GAR TFase; EC 2.1.2.2) has been purified 70-fold to apparent homogeneity from Escherichia coli harboring an expression vector encoding the purN gene product, GAR TFase. The protein is a monomer of Mr 23,241 and catalyzes a single reaction. Steady-state ki ... >> More

  Glycinamide ribonucleotide transformylase (GAR TFase; EC 2.1.2.2) has been purified 70-fold to apparent homogeneity from Escherichia coli harboring an expression vector encoding the purN gene product, GAR TFase. The protein is a monomer of Mr 23,241 and catalyzes a single reaction. Steady-state kinetic parameters for the enzyme have been obtained. The structural requirements for cofactor utilization have been investigated and found to parallel those of the multifunctional avian enzyme. The enzyme was inactivated with the affinity label N10-(bromoacetyl)-5,8-dideazafolate in a stoichiometric and active-site-specific manner. The ionization state of the cofactor analogue in the enzyme-cofactor complex appears to require the dissociation of the proton at N3 of the pyrimidine within the complex. << Less

  Biochemistry 29:1436-1443(1990) [PubMed] [EuropePMC]

• N10-Formyltetrahydrofolate is the formyl donor for glycinamide ribotide transformylase in Escherichia coli.

  Dev I.K., Harvey R.J.

  Glycinamide ribotide transformylase from Escherichia coli was obtained free of N5,N10-methenyltetrahydrofolate cyclohydrolase activity by DEAE-cellulose chromatography. In reaction mixtures containing this enzyme preparation in potassium maleate buffer, pH 7.2, no detectable interconversion of N5, ... >> More

  Glycinamide ribotide transformylase from Escherichia coli was obtained free of N5,N10-methenyltetrahydrofolate cyclohydrolase activity by DEAE-cellulose chromatography. In reaction mixtures containing this enzyme preparation in potassium maleate buffer, pH 7.2, no detectable interconversion of N5,N10-methenyltetrahydrofolate occurred. Upon addition of glycinamide ribotide, N-formylglycinamide ribotide was formed when N10-formyltetrahydrofolate was present; no formylation occurred in the presence of N5,N10-methenyltetrahydrofolate. A method for the synthesis and purification of glycinamide ribotide is presented. << Less

  J. Biol. Chem. 253:4242-4244(1978) [PubMed] [EuropePMC]

• L(-)-10-Formyltetrahydrofolate is the cofactor for glycinamide ribonucleotide transformylase from chicken liver.

  Smith G.K., Benkovic P.A., Benkovic S.J.

  It is shown that L(-)-10-formyltetrahydrofolate serves as the cofactor for glycinamide ribonucleotide transformylase from chicken liver. The utilization of L(-)-10-formyl-H4folate was not previously recognized, because L-(+)-10-formyl-H4folate is an excellent competitive inhibitor of the enzyme, K ... >> More

  It is shown that L(-)-10-formyltetrahydrofolate serves as the cofactor for glycinamide ribonucleotide transformylase from chicken liver. The utilization of L(-)-10-formyl-H4folate was not previously recognized, because L-(+)-10-formyl-H4folate is an excellent competitive inhibitor of the enzyme, Ki = 0.75 +/-0.07 microM, and historically the transformylase assay was carried out with a mixture of diastereomers. The results are discussed in relation to the utilization of L(+)-5,10-methenyl-H4folate. << Less

  Biochemistry 20:4034-4036(1981) [PubMed] [EuropePMC]

• Biosynthesis of the purines. XIX. 2-Amino-N-ribosylacetamide 5'-phosphate (glycinamide ribotide) transformylase.

  WARREN L., BUCHANAN J.M.

  J Biol Chem 229:613-626(1957) [PubMed] [EuropePMC]

• Biosynthesis of the purines. XXVI. The identification of the formyl donors of the transformylation reactions.

  HARTMAN S.C., BUCHANAN J.M.

  J Biol Chem 234:1812-1816(1959) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Human glycinamide ribonucleotide transformylase: active site mutants as mechanistic probes.

  Manieri W., Moore M.E., Soellner M.B., Tsang P., Caperelli C.A.

  Human glycinamide ribonucleotide transformylase (GART) (EC 2.1.2.2) is a validated target for cancer chemotherapy, but mechanistic studies of this therapeutically important enzyme are limited. Site-directed mutagenesis, initial velocity studies, pH-rate studies, and substrate binding studies have ... >> More

  Human glycinamide ribonucleotide transformylase (GART) (EC 2.1.2.2) is a validated target for cancer chemotherapy, but mechanistic studies of this therapeutically important enzyme are limited. Site-directed mutagenesis, initial velocity studies, pH-rate studies, and substrate binding studies have been employed to probe the role of the strictly conserved active site residues, N106, H108, and D144, and the semiconserved K170 in substrate binding and catalysis. Only two conservative substitutions, N106Q and K170R, resulted in catalytically active enzymes, and these active mutant enzymes gave pH-rate profiles and a steady-state kinetic mechanism essentially identical to those of the native enzyme. All inactive mutants were able to bind both substrates, ruling out disrupted formation of the ternary complex as the source of inactivity. Differences between human and Escherichia coli GART, previously used as a model for the human enzyme, were evident. << Less

  Biochemistry 46:156-163(2007) [PubMed] [EuropePMC]