Properties of thymidylate synthetase from dichloromethotrexate-resistant Lactobacillus casei.

Abstract Extracts of dichloromethotrexate-resistant Lactobacillis casei provide a rich source of stable thymidylate synthetase, showing 100 times the activity found in extracts of the sensitive strain. The enzyme was purified 17-fold from extracts of resistant cells. It has a molecular weight of 67,000 and is activated by 2-mercaptoethanol. The rate of the reaction as determined by loss of tritium from deoxyuridine monophosphate-5-3H was 84% of the rate observed spectrophotometrically. The enzyme from dichloromethotrexate-resistant L. casei differs from Escherichia coli thymidylate synthetase, in that it will utilize tetrahydrohomofolate as a cofactor in place of tetrahydrofolate. The Km values for tetrahydrofolate and tetrahydrohomofolate are 4.5 x 10-5 m and 7.2 x 10-6 m, respectively, showing that tetrahydrohomofolate is bound to the enzyme more tightly than tetrahydrofolate, although the Vmax obtained with tetrahydrohomofolate is only 6% of that obtained with tetrahydrofolate. The Ki of tetrahydrohomofolate of 4.6 x 10-6 m is close to the Km for that compound. 10-Methyltetrahydrofolate and 11-methyltetrahydrohomofolate were inactive as cofactors. They inhibited the reaction 50% at 4 x 10-6 m and 3 x 10-5 m, respectively. Tetrahydrohomopteroate was neither a substrate nor an inhibitor at 3 x 10-4 m. Methotrexate-resistant strains of L. casei also showed high levels of thymidylate synthetase activity.

• Publication year

  1970

• Venue

  Journal of Biological Chemistry

• Publication date

  1970-10-25

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1016/s0021-9258(18)62753-xPMID 4918842
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Studies with tetrahydrohomofolate and thymidylate synthetase from amethopterin-resistant mouse leukemia cells.

  1968cited by this paper
• The toxicity of tetrahydrohomofolic acid in normal and malaria-infected monkeys.

  1968cited by this paper
• A new assay of thymidylate synthetase activity based on the release of tritium from deoxyuridylate-5-3-H.

  1967cited by this paper
• The purification and properties of thymidylate synthetase from chick embryo extracts.

  1967cited by this paper
• Antimalarial Activity of Tetrahydrohomopteroic Acid

  1967cited by this paper
• Folate coenzymes in amethopterin-sensitive and -resistant strains of Streptococcus faecalis. Enzymatic formation and metabolic function.

  1966cited by this paper
• A PURIFICATION OF BREWERS' AND BAKERS' YEAST ENOLASE YIELDING A SINGLE ACTIVE COMPONENT.

  1964cited by this paper
• The kinetics of enzyme-catalyzed reactions with two or more substrates or products. II. Inhibition: nomenclature and theory.

  1963cited by this paper
• The association of increased dihydrofolate reductase with amethopterin resistance in mouse leukemia.

  1962cited by this paper
• The enzymatic synthesis of thymidylate. I. Early steps in the purification of thymidylate synthetase of Escherichia coli.

  1962cited by this paper
• The enzymatic synthesis of thymidylate. III. The further purification of thymidylate synthetase and its separation from natural fluorescent inhibitors.

  1962cited by this paper
• Protein measurement with the Folin phenol reagent.

  1951cited by this paper

• Comparative study of Nucleic Acid anabolism pathway in Medical Leech Hirudo medicinals and Earthworm Aporrectoda trapezoides

  2019cites this paper
• Inhibition of thymidylate synthase by the diastereoisomers of leucovorin

  2008cites this paper
• Shotgun and Characterization of the Thymidylate Synthase-Encoding Gene from Mycobacterium bovis BCG

  2007cites this paper
• Yoshida into DNA of HL-60 Cells by Metabolic Modulations Enhancement of the Incorporation of 5-Fluorodeoxyuridylate Updated Version

  2006cites this paper
• Competitive BindingRadioassayfor 5-Fluorodeoxyuridine 5'-.Monophosphatein Tissues

  2006cites this paper
• Effects of Mg2+ and adenine nucleotides on thymidylate synthetase from different mouse tumors

  2004cites this paper
• Economizing DNA-specific labelling by deoxythymidine-5′-monophosphate in Saccharomyces cerevisiae

  2004cites this paper
• Bacteriophage Resistance of a ΔthyA Mutant of Lactococcus lactis Blocked in DNA Replication

  2002cites this paper
• Inhibition of thymidylate synthase by pergularinine, tylophorinidine and deoxytubulosine.

  1999cites this paper
• Shotgun Cloning and Characterization of the Thymidylate Synthase‐Encoding Gene from Mycobacterium bovis BCG

  1998cites this paper
• The targeting of factors necessary for expression of methicillin resistance in staphylococci.

  1998cites this paper
• beta-Carboline-benzoquinolizidine alkaloid deoxytubulosine inhibits thymidylate synthase activity in leukemic leukocytes from patients with chronic myeloblastic leukemia and acute lymphoblastic leukemia.

  1998cites this paper
• Over-expression of target genes as a mechanism of antibiotic resistance in bacteria.

  1998cites this paper
• Thymidylate synthase activity in leukocytes from patients with chronic myelocytic leukemia and acute lymphocytic leukemia and its inhibition by phenanthroindolizidine alkaloids pergularinine and tylophorinidine.

  1998cites this paper
• Inhibition of thymidylate synthase and cell growth by the phenanthroindolizidine alkaloids pergularinine and tylophorinidine.

  1997cites this paper
• The catalytic mechanism and structure of thymidylate synthase.

  1995cites this paper
• Physico-chemical properties of thymidylate synthase from Lactobacillus leichmannii: thiol groups, amino acid composition and the terminal end-groups.

  1994cites this paper
• Purification, relative molecular mass and subunits of thymidylate synthase from Lactobacillus leichmannii.

  1993cites this paper
• Refined structures of substrate-bound and phosphate-bound thymidylate synthase from Lactobacillus casei.

  1993cites this paper
• Ternary complex formation with 5-fluoro-2'-deoxyuridylate and the kinetic properties of thymidylate synthase from Lactobacillus leichmannii.

  1993cites this paper
• Purine and Pyrimidine Salvage Pathways

  1993cites this paper
• Characteristics of NADPH-dependent thymidylate synthetase purified from Streptomyces aureofaciens.

  1992cites this paper
• Folate analogues. 35. Synthesis and biological evaluation of 1-deaza, 3-deaza, and bridge-elongated analogues of N10-propargyl-5,8-dideazafolic acid.

  1991cites this paper
• Tetrahydrohomofolate polyglutamates as inhibitors of thymidylate synthase and glycinamide ribonucleotide formyltransferase in Lactobacillus casei.

  1990cites this paper
• Cloning and characterization of the thymidylate synthase gene from Lactococcus lactis subsp. lactis

  1990cites this paper
• Recent Studies on Thymidylate Synthase

  1989cites this paper
• Hydride transfer by dihydrofolate reductase. Causes and consequences of the wide range of rates exhibited by bacterial and vertebrate enzymes.

  1989cites this paper
• Expression of human thymidylate synthase in Escherichia coli.

  1989cites this paper
• Biomedical science and the third world. Under the volcano. Recent studies on thymidylate synthase.

  1989cites this paper
• and Sustained Release of Its Active Metabolites in VX-2 Rabbit Hepatoma followingIntraarterial Administration of FdUrd-C8 Solution in Lipiodol

  1988cites this paper
• Overview: rational basis for development of fluoropyrimidine/5-formyltetrahydrofolate combination chemotherapy.

  1988cites this paper
• Synthesis and biological evaluation of 8-deazahomofolic acid and its tetrahydro derivative.

  1988cites this paper
• Selective accumulation of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8) and sustained release of its active metabolites in VX-2 rabbit hepatoma following intraarterial administration of FdUrd-C8 solution in Lipiodol.

  1988cites this paper
• Cloning, Sequencing, and Expression of the Lactobacillus casei Thymidylate Synthase Gene

  1988influential citation
• Kinetics and thermodynamics of the interaction of 5-fluoro-2'-deoxyuridylate with thymidylate synthase.

  1987cites this paper
• Atomic structure of thymidylate synthase: target for rational drug design.

  1987cites this paper
• Folate Biochemistry and the Metabolism of One-Carbon Units

  1987cites this paper
• Kinetics and mechanism of interaction of 10-propargyl-5,8-dideazafolate with thymidylate synthase.

  1986cites this paper
• Preparation of (6R)-tetrahydrofolic acid and (6R)-5-formyltetrahydrofolic acid of high stereochemical purity.

  1986cites this paper
• Cytotoxic effects of folate antagonists against methotrexate-resistant human leukemic lymphoblast CCRF-CEM cell lines.

  1985cites this paper
• Purification and characterization of the bifunctional thymidylate synthetase-dihydrofolate reductase from methotrexate-resistant Leishmania tropica.

  1985cites this paper
• Molecularand Karyological Analysisof Methotrexate-resistant and -sensitive HumanLeukemicCCRF-CEMCells1

  1985cites this paper
• Thymidylate Synthetase Inhibitors: Experimental and Clinical Aspects

  1985cites this paper
• Coregulation of dihydrofolate reductase and thymidylate synthase B in bacillus subtilis.

  1985cites this paper
• Molecular and karyological analysis of methotrexate-resistant and -sensitive human leukemic CCRF-CEM cells.

  1985cites this paper
• Capabilities of microorganisms (and microbiologists).

  1984cites this paper
• Interaction of N4-hydroxy-2'-deoxycytidylic acid with thymidylate synthetase.

  1984cites this paper
• Enhancement of the incorporation of 5-fluorodeoxyuridylate into DNA of HL-60 cells by metabolic modulations.

  1983cites this paper
• The interaction of pteroylpolyglutamates with calf thymus thymidylate synthase.

  1983cites this paper
• Association of RNA with thymidylate synthase from methotrexate-resistant Streptococcus faecium.

  1983cites this paper
• An hypothesis on the role of pteroylpolyglutamate derivatives as coenzymes.

  1983cites this paper
• Crystal structures of Escherichia coli and Lactobacillus casei dihydrofolate reductase refined at 1.7 A resolution. I. General features and binding of methotrexate.

  1982cites this paper
• Secondary alpha-hydrogen isotope effects on the interaction of 5-fluoro-2'-deoxyuridylate and 5,10-methylenetetrahydrofolate with thymidylate synthetase.

  1982cites this paper
• Properties of thymidylate synthetase from Ehrlich ascites carcinoma cells. Effect of Mg2/ and MgATP2-.

  1982cites this paper
• Purification of mouse liver thymidylate synthetase by affinity chromatography using 10-methyl-5,8-dideazafolate as the affinant

  1981cites this paper
• Polyglutamyl derivatives of tetrahydrofolate as substrates for Lactobacillus casei thymidylate synthase.

  1981cites this paper
• Inhibition of serine metabolism by tetrahydrohomofolate in L1210 mouse leukemia cells.

  1981cites this paper
• Conservation and variation of nucleotide sequences within related bacterial genomes: Escherichia coli strains

  1980cites this paper
• Biochemical and genetic basis of the response to 5-fluorouracil in Drosophila melanogaster

  1980cites this paper
• Interaction of thymidylate synthetase with 5-nitro-2'-deoxyuridylate.

  1980cites this paper
• [6] Use of secondary α-hydrogen isotope effects in reactions of pyrimidine nucleoside/nucleotide metabolism: Thymidylate synthetase

  1980cites this paper
• Synthesis of 5-[(methylthio)methyl]-2'-deoxyuridine, the corresponding sulfoxide and sulfone, and their 5'-phosphates: antiviral effects and thymidylate synthetase inhibition.

  1980cites this paper
• A calorimetric study of the binding of 2'-deoxyuridine-5'-phosphate and its analogs to thymidylate synthetase.

  1980cites this paper
• Folate analogues altered in the C9-N10 bridge region: 11-thiohomofolic acid.

  1979cites this paper
• Thymidylate synthase activity from chlamydomonas cells and cultured tissues of Nicotiana, pinus, and daucus.

  1979cites this paper
• 31P NMR studies on the interaction of deoxyuridylate with thymidylate synthase.

  1979cites this paper
• Enzyme affinity of the 5,6-dihydro derivatives of the substrate and product of thymidylate synthetase catalysis.

  1979cites this paper
• A biochemical and pharmacological study of therapeutic synergism with 5-fluorouracil plus cyclophosphamide in murine L1210 leukemia.

  1979cites this paper
• Thymidylate synthetase. Catalysis of dehalogenation of 5-bromo- and 5-iodo-2'-deoxyuridylate.

  1979cites this paper
• 5-[(4-Methyl-1,2,3,4-tetrahydroquinoxalyl)methyl]-2'-deoxyuridine 5'-phosphate: an analogue of a proposed intermediate in thymidylate synthetase catalysis.

  1979cites this paper
• Mechanisms and distribution of bacterial resistance to diaminopyrimidines and sulphonamides.

  1979cites this paper
• Thymidylate synthetase activity in normal and brugia pahangi-infected aedes aegypti.

  1979cites this paper
• Inhibition of thymidylate synthetase of the human lymphocytes.

  1979cites this paper
• Competitive binding radioassay for 5-fluorodeoxyuridine 5'-monophosphate in tissues.

  1979cites this paper
• 5-Cyano-2'-deoxyuridine 5'-phosphate: a potent competitive inhibitor of thymidylate synthetase.

  1979cites this paper
• Thymidylate synthetase from Escherichia coli K12. Purification, and dependence of kinetic properties on sugar conformation and size of the 2' substituent.

  1979cites this paper
• Thymidylate synthetase catalyzed exchange of tritiumfrom [5-3H]-2'-deoxyuridylate for protons of water.

  1979cites this paper
• Mechanism of resistance of Pediococcus cerevisiae strains to 5-fluorodeoxyuridine.

  1978cites this paper
• Thymidylate synthetase and dihydrofolic acid reductase in the stimulated human lymphocyte

  1978cites this paper
• Dihydrofolate reductase from Lactobacillus casei. X-ray structure of the enzyme methotrexate.NADPH complex.

  1978cites this paper
• Binding of N5,N10-methylene tetrahydrofolate and the inhibition of thymidylate synthesis by a folate-binding protein.

  1978cites this paper
• Effects of 5-fluorouracil on 5-fluorodeoxyuridine 5'-monophosphate and 2-deoxyuridine 5'-monophosphate pools, and DNA synthesis in solid mouse L1210 and rat Walker 256 tumors.

  1978cites this paper
• 5-Nitro-2'-deoxyuridine 5'-monophosphate is a potent irreversible inhibitor of Lactobacillus caesi thymidylate synthetase.

  1978cites this paper
• 5-Nitro-2'-deoxyuridylate: a mechanism-based inhibitor of thymidylate synthetase.

  1978cites this paper
• Effects of 5-fluorouracil on human colon carcinoma and solid rat Walker 256 carcinosarcoma: evaluation as in vitro predictors of clinical response.

  1978cites this paper
• Tumor 5-fluorodeoxyuridylate concentration as a determinant of 5-fluorouracil response.

  1978cites this paper
• Inhibition of Lactobacillus casei thymidylate synthetase by 5-substituted 2'-deoxyuridylates. Preliminary quantitative structure-activity relationship.

  1977cites this paper
• A calorimetric study of the binding of 2'-deoxyuridine-5'-phosphate and 5-fluoro-2'-deoxyuridine-5'-phosphate to thymidylate synthetase.

  1977cites this paper
• Understanding the actions of carcinostatic drugs to improve chemotherapy: 5-fluorouracil.

  1977cites this paper
• Complex inhibition of thymidylate synthetase by aromatic diamidines: evidence for both rapid, freely reversible and slowly progressive, nonequilibrium inhibition.

  1977cites this paper
• Reduction of oxidised folates by dihydrofolate reductase from methotrexate-resistant Lactobacillus casei.

  1977cites this paper
• [Thymidylate synthetase (author's transl)].

  1977cites this paper
• Increased Thymidylate Synthetase Activity in 5-Fluorodeoxyuridine-Resistant Novikoff Hepatoma Cells 1

  1977cites this paper
• Presence and properties of thymidylate synthase in trypanosomatids.

  1977cites this paper
• Active-site labeling of thymidylate synthetase with 5-fluoro-2'-deoxyuridylate.

  1977cites this paper
• Rate-limiting steps in folate metabolism by Lactobacillus casei.

  1977cites this paper
• Thymidylate synthetase: studies on peptide containing covalently bound 5-fluoro-2'-deoxyuridylate and 5,10-methylenetetrahydrofolate.

  1976cites this paper
• Formation of ternary complexes of thymidylate synthetase as followed by absorbance, fluorescence, and circular dichroic spectra and gel electrophoresis.

  1976cites this paper
• Purification of thymidylate synthetase from enzyme-poor sources by affinity chromatography.

  1976cites this paper
• Thymidylate synthetase: fluorine-19 nmr characterization of the active site peptide covalently bound to 5-fluoro-2'-deoxyuridylate and 5,10-methylenetetrahydrofolate.

  1976cites this paper