Wyniki wyszukiwania

1

Oczyszczanie enzymów i innych białek biologicznie czynnych poprzez chromatografię na substancjach niskocząsteczkowych unieruchomionych w fazie stałej

100%

Rode W.

Postępy Biochemii

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1977

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tom 23

|

nr 1

2

Biosynteza tymidylanu: rola biologiczna i regulacja w komórkach zwierzęcych

100%

Rode W.

Postępy Biochemii

|

1986

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tom 32

|

nr 4

3

Inhibicja syntazy tymidylanowej w chemioterapii

100%

Rode W.

Postępy Biochemii

|

1996

|

tom 42

|

nr 2

133-140

4

Specifity of thymidylate synthase inactivation by 4,5-bisubstituted dUMP analogues

100%

Rode W.

Acta Biochimica Polonica

|

1993

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tom 40

|

nr 3

5

Syntetaza tymidylanowa

75%

Rode W.

Postępy Biochemii

|

1977

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tom 23

|

nr 1

6

Methionine synthase assay based on coupling whit thymidylate synthase reaction

63%

Rode W., Balinska M.

Acta Biochimica Polonica

|

1985

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tom 32

|

nr 2

7

An isotopic assay of dUTPase activity based on coupling with thymidylate synthase

63%

Golos B., Rode W.

Acta Biochimica Polonica

|

1999

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tom 46

|

nr 3

A new rapid, sensitive and convenient procedure is presented allowing determination of dUTPase activity. With [5-3H]dUTP used as the substrate, dUTPase, converts it to the corresponding monophosphate and is coupled with a thymidylate synthase-catalyzed reaction, resulting in tritium release from [5-3H]dUMP. Following charcoal absorption of the labeled nuleotides, radioactivity of tritiated water is determined. The new assay was tested to show comparable results with a previously described assay, based on measuring dUTPase-catalyzed [5-3H]dUMP production.

8

Molecular mechanism of thymidylate synthase-catalysed reaction and interaction of the enzyme with 2- and-or 4-substituted analogues of dUMP and 5-fluoro-dUMP

63%

Rode W., Les A.

Acta Biochimica Polonica

|

1996

|

tom 43

|

nr 1

Thymidylate synthase is a target enzyme in anticancer, antiviral, antifungal and antiprotozoan chemotherapy. With two dUMP analogues, 5-fluoro-dUMP (FdUMP) and 5-(trifluoromethyl)-dUMP (CF3dUMP), strong thymidylate synthase inhibitors and active forms of drugs, the inhibition mechanism is based on the reaction mechanism. Recent comparative studies of new dUMP analogues, containing more than one substituent in the pyrimidine ring, showed that substitution of the pyrimidine ring Qd=0 group in FdUMP by either C(4)=N-OH group (in N4-hydroxy-FdCMP) or C(4)=S group (in 4-thio-FdUMP) preserves high inhibitory potency of the drug but may alter its specificity for thymidylate synthases from various sources, which differ in sensitivity to slow-binding inhibition by FdUMP. Informations suggesting mechanisms responsible for the foregoing have been reviewed, including results of molecular modeling studies suggesting interaction of the pyrimidine C(4)=0 group, or its modification, with the N4-methylene-

9

Themidylate synthase activity in the development of Hymenolepis diminuta

51%

Ciesla J., Zielinski Z., Machnicka B., Rode W.

Acta Biochimica Polonica

|

1987

|

tom 34

|

nr 3

10

Phosphorylation of basic amino acid residues in proteins: important but easily missed

51%

Ciesla J., Fraczyk T., Rode W.

Acta Biochimica Polonica

|

2011

|

tom 58

|

nr 2

Reversible phosphorylation is the most widespread posttranslational protein modification, playing regulatory role in almost every aspect of cell life. The majority of protein phosphorylation research has been focused on serine, threonine and tyrosine that form acid-stable phosphomonoesters. However, protein histidine, arginine and lysine residues also may undergo phosphorylation to yield acid-labile phosphoramidates, most often remaining undetected in conventional studies of protein phosphorylation. It has become increasingly evident that acid-labile protein phosphorylations play important roles in signal transduction and other regulatory processes. Beside acting as high-energy intermediates in the transfer of the phosphoryl group from donor to acceptor molecules, phosphohistidines have been found so far in histone H4, heterotrimeric G proteins, ion channel KCa3.1, annexin 1, P-selectin and myelin basic protein, as well as in recombinant thymidylate synthase expressed in bacterial cells. Phosphoarginines occur in histone H3, myelin basic protein and capsidic protein VP12 of granulosis virus, whereas phospholysine in histone H1. This overview of the current knowledge on phosphorylation of protein basic amino-acid residues takes into consideration its proved or possible roles in cell functioning. Specific requirements of studies on acid-labile protein phosphorylation are also indicated.

11

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Porownanie wlasciwosci syntazy tymidylanowej oczyszczonej z tasiemca szczurzego, Hymenolepis diminuta, z wlasciwosciami enzymu zywiciela wyizolowanego z regenerujacej watraby szczura

45%

Ciesla J., Machnicka B., Pawelczak K., Rzeszotarska B., Rode W.

Wiadomości Parazytologiczne

|

1992

|

tom 38

|

nr 1-2

23-29

Thymidylate synthases (TS) from the tapeworm, Hymenolepis diminuta, and regenerating rat liver have been purified by means of affinity chromatography on immobilized 10-formyl-5,8-dideazafolate and concentrated on immobilized p-aminophenyl-5-fluoro-2'-deoxyuridine monophosphate. Molecular weights of native TS from the tapeworm and regenerating rat liver were 62 kD and 81.5 kD, respectively, and molecular weights of the monomers were 34.4 kD and 34.9 kD, respectively, painting to dimeric structures of both enzymes. The dependence of TS activity on temperature (ARRHENIUS plot) was biphasic for the parasite enzyme, with lower activation energy above 32°C, and monophasic for the host enzyme. 2'-deoxyuridine-5'-monophosphate (dUMP) analogues, 5-fluoro-2'-deoxyuridine-5' -monophosphate (5-FdUMP), 2-tio-5-FdUMP, N⁴-hydroxy-2'-deoxycytidine-5'-monophosphate (N⁴-hydroxy-dCMP) and N⁴-hydroxy-5-FdCMP, were competitive with respect to dUMP, slow-binding inhibitors of TS from both sources, with K₁ values in 10⁻⁶ - 10⁻⁹ M range. 5-FdUMP was distinctly stronger inhibitor of the host than the tapeworm TS, whereas N⁴-hydroksy-5-FdCMP inhibited stronger the parasite enzyme. Interactions of 5,10-methylenetetrahydrofolate (CH₂H₄PteGlu) analogue, 10-propargyl-5,8-dideazafolate (pddPteGlu), and its di- and triglutamates with both enzymes were studied. Inhibition of the parasite and host enzymes by pddPteGlu was of mixed-type with respect to CH₂H₄PteGlu, with K₁ values in 10⁻⁸ M range. Introduction of additional glutamate residues changed inhibition type to noncompetitive with respect to CH₂H₄PteGlu and lowered K₁ values (pddPteGlu₃ < pddPteGlu₂ < pddPteGlu₁). The latter potentiation of inhibitory properties was distinctly stronger in case of the tapeworm than regenerating rat liver TS.

12

Altered mouse leukemia L1210 thymidylate synthase, associated with cell resistance to 5-fluoro-dUrd, is not mutated but rather reflects posttranslational modification

45%

Ciesla J., Fraczyk T., Zielinski Z., Sikora J., Rode W.

Acta Biochimica Polonica

|

2006

|

tom 53

|

nr 1

Thymidylate synthase purified from 5-fluoro-dUrd-resistant mouse leukemia L1210 cells (TSr) was less sensitive to slow-binding inhibition by 5-fluoro-dUMP than the enzyme from the parental cells (TSp), both enzyme forms differing also in sensitivity to several other dump analogues, apparent molecular weights of monomer and dimer, and temperature dependence of the catalyzed reaction. Direct sequencing of products obtained from RT-PCR, performed on total RNA isolated from the parental and 5-fluoro-dUrd-resistant cells, proved both nucleotide sequences to be identical to the mouse thymidylate synthase coding sequence published earlier (NCBI protein database access no. NP_067263). This suggests that the altered properties of TSr are caused by a factor different than protein mutation, presumably posttranslational modification. As a possibility of rat thymidylate synthase phosphorylation has been recently demonstrated (Samsonoff et al. (1997) J Biol Chem 272: 13281), the mouse enzyme amino-acid sequence was analysed, revealing several potential phosphorylation sites. In order to test possible influence of the protein phosphorylation state on enzymatic properties, endogenous TSp and TSr were purified in the presence of inhibitors of phosphatases. Although both enzyme forms were phosphorylated, as shown by electrophoretical separation followed by phosphoprotein detection, the extent of phosphorylation was apparently similar. However, the same two purified enzyme preparations, compared to the corresponding preparations purified in the absence of phosphatase inhibitors, showed certain properties, including sensitivity to the slow-binding inhibition by FdUMP, altered. Thus properties dependence on phosphorylation was indicated.

13

Thymidylate biosynthesis as a target in chemotherapy. Potential importance of a peculiar expression pattern of enzymes involved in thymidylate biosynthesis in nematodes

39%

Rode W., Dabrowska M., Golos B., Winska P., Jagielska E., Ciesla J., Zielinski Z.

Acta Biochimica Polonica

|

2003

|

tom 50

|

nr Supplement 1

14

Comparison of some diaminopyrimidines and amethopterin as inhibitors of insect cell proliferation and of insect folate-metabolizing enzymes

39%

Chmurzynska W., Grzelakowska-Sztabert B., Manteuffel-Cymborowska M., Rode W., Saska J., Zielinska Z. M.

Acta Biochimica Polonica

|

1974

|

tom 21

|

nr 4

15

The effect of Arg209 to Lys mutation in mouse thymidylate synthase

39%

Ciesla J., Golos B., Walajtys-Rode E., Jagielska E., Plucienniczak A., Rode W.

Acta Biochimica Polonica

|

2002

|

tom 49

|

nr 3

Mouse thymidylate synthase R209K (a mutation corresponding to R218K in Lactobacillus casei), overexpressed in thymidylate synthase-deficient Escherichia coli strain, was poorly soluble and with only feeble enzyme activity. The mutated protein, incubated with FdUMP and N5,10-methylenetetrahydrofolate, did not form a complex stable under conditions of SDS/polyacrylamide gel electrophoresis. The reaction catalyzed by the R209K enzyme (studied in a crude extract), compared to that catalyzed by purified wild-type recombinant mouse thymidylate synthase, showed the Km value for dUMP 571-fold higher and Vmax value over 50-fold (assuming that the mutated enzyme constituted 20% of total crude extract protein) lower. Thus the ratios kcat,R209K/kcat,'wild' and (kcat, R209K/Km, R209K dUMP)/ (kcat, 'wild'/Km, 'wild' dUMP) were 0.019 and 0.000032, respectively, documenting that mouse thymidylate synthase R209, similar to the corresponding L. casei R218, is essential for both dUMP binding and enzyme reaction.

16

Sulfamide antifolates inhibiting thymidylate synthase: synthesis, enzyme inhibition and cytotoxicity

39%

Pawelczak K., Makowski M., Kempny M., Dzik J. M., Golos B., Rode W., Rzeszotarska B.

Acta Biochimica Polonica

|

2002

|

tom 49

|

nr 2

Synthesis and biological evaluation are described of seven new analogues (3-9) of two potent thymidylate synthase inhibitors, 10-propargyl-5,8-dideazafolate (1) and its 2-methyl-2-deamino congener ICI 198583 (2). While the new compunds 3 and 4 were analogues of 1 and 2, respectively, containing a p-aminobenzenesulfonyl residue in place of the p-aminobenzoic acid residue, the remaining 5 new compounds were analogues of 4 with the L-glutamic acid residue replaced by glycine (5), L-valine (6), L-alanine (7), L-phenylglycine (8) or L-norvaline (9). The new analogues were tested as inhibitors of thymidylate synthases isolated from tumour (Ehrlich carcinoma), parasite (Hymenolepis diminuta) and normal tissue (regenerating rat liver) and found to be weaker inhibitors than the parent 10-propargyl-5,8-dideazafolic acid. Selected new analogues, tested as inhibitors of growth of mouse leukemia L 5178Y cells, were less potent than the parent 10-propargyl-5,8-dideazafolic acid. Substitution of the glutamyl residue in compound 4 with L-norvaline (9) resulted in only a 5-fold stronger thymidylate synthase inhibitor, but a 40-fold weaker cell growth inhibitor.

17

Acyclic analogues of 5-fluoro-dUMP and 5-fluoro-2'-deoxyuridine: Synthesis and inhibition of thymidylate synthase and tumour cell growth

39%

Felczak K., Golos B., Dzik J., Rode W., Bretner M., Shugar D., Kulikowski T.

Acta Biochimica Polonica

|

1998

|

tom 45

|

nr 1

1-[(2-Hydroxyethoxy)methyl]-5-fluorouracil (HEMFU) and 1-[(1,3-dihydroxy-2-propoxy)methyl]-5-fluorouracil (DHPFU) were prepared by alkylation of the di-O-TMS derivative of 5-fluorouracil and phosphorylated with the use of the wheat shoot phosphotransferase system to their monophosphates, HEMFUMP and DHPFUMP. 1-(2-Phosphonylmethoxyethyl)-5-fluorouracil (PMEFU) was obtained by condensation of diethyl-2-chloroethoxymethanephosphonate with 5-fluorouracil and cleavage of the alkylphosphoester with trimethylbromosilane. Inhibition of highly purified thymidylate synthase from mouse tumour Ehrlich carcinoma and leukemia L1210 cells by each of the nucleotide analogues, DHPFUMP, PMEFU and HEMFUMP, and of L5178Y mouse leukemia cell growth by the nucleoside (HEMFU) analogue, were studied. DHPFUMP proved to be the strongest inhibitor, non-competitive vs dUMP, with Kiapp 2.8 μM for time-independent interaction with the enzyme and N5,N10-methylenetetrahydrofolate (CH2H4PteGlu). In the presence of CH2H4PteGlu, DHPFUMP exhibited time-dependent inactivation of the enzyme, the inactivation rate plots being biphasic and pointing to Ki values in the μM range (103-fold higher than for 5-fluoro-dUMP). HEMFUMP and PMEFU were much weaker inhibitors of the enzyme, with Kiapp values of 0.26 μM (non-competitive vs dUMP) and 30 mM (non-competitive vs dUMP), respectively. HEMFU, despite the weak interaction of its nucleotide analogue with the enzyme, proved to be a strong cell (L5178Y) growth inhibitor, with IC50 in the range 10-5 M.

18

Synthesis and interaction with thymidylate synthase of 5'-dithiophosphate and 5'-fluorothiophosphate of thymidine

39%

Golos B., Misiura K., Olesiak M., Okruszek A., Stec W. J., Rode W.

Acta Biochimica Polonica

|

1998

|

tom 45

|

nr 1

Thymidine-5'-fluorothiophosphate, dTMP(S)-F, was synthesized by the oxathiaphospholane, and thymidine 5'-dithiophosphate, dTMPS2, by the dithiaphospholane, method. To estimate the role of 5'-phosphate group ionization in binding of pyrimidine nucleotides by thymidylate synthase, dTMP(S)-F was studied as an inhibitor of mouse tumour (L1210) enzyme, and its inhibitory properties were compared with those of dTMPS2, a close dTMP analogue. While dTMPS2 proved to be an inhibitor, competitive vs dUMP, with Kiapp = 94 μM, the 5'-fluorothiophosphate congener displayed no activity, indicating that the enzyme requires for binding the presence of a dianionic 5'-phosphate group in a nucleotide.

19

Synthesis and biological activity of Na-[4-[N-[[3,4-dihydro-2-methyl-4-oxo-6-quinazolinyl]methyl]-N-propargylamino]phenylacetyl]-L-glutamic acid

33%

Kusakiewicz-Dawid A., Bugaj M., Dzik J. M., Golos B., Winska P., Pawelczak K., Rzeszotarska B., Rode W.

Acta Biochimica Polonica

|

2002

|

tom 49

|

nr 1

2-Deamino-2-methyl-N10-propargyl-5,8-dideazafolic acid (ICI 198583) is a potent inhibitor of thymidylate synthase. Its analogue, Nα-[4-[N-[(3,4-dihydro 2-methyl-4 -oxo- 6-quinazolinyl)methyl]-N-propargylamino]phenylacetyl]-L-glutamic acid, containing p-aminophenylacetic acid residue substituting p-aminobenzoic acid residue, was synthesized. The new analogue exhibited a moderately potent thymidylate synthase inhibition, of linear mixed type vs. the cofactor, N5,10 -methylenetetrahydrofolate. The K value of 0.34 uM, determined with a purified recombinant rat hepatoma enzyme, was about 30-fold higher than that reported for inhibition of thymidylate synthase from mouse leukemia L1210 cells by ICI 198583 (Hughes et al., 1990, J. Med. Chem. 33, 3060). Growth of mouse leukemia L5178Y cells was inhibited by the analogue (IC50 = 1.26 mM) 180-fold weaker than by ICI 198583 (IC50 = 6.9 uM).

Ograniczanie wyników

Oczyszczanie enzymów i innych białek biologicznie czynnych poprzez chromatografię na substancjach niskocząsteczkowych unieruchomionych w fazie stałej

Biosynteza tymidylanu: rola biologiczna i regulacja w komórkach zwierzęcych

Inhibicja syntazy tymidylanowej w chemioterapii

Specifity of thymidylate synthase inactivation by 4,5-bisubstituted dUMP analogues

Syntetaza tymidylanowa

Methionine synthase assay based on coupling whit thymidylate synthase reaction

An isotopic assay of dUTPase activity based on coupling with thymidylate synthase

Molecular mechanism of thymidylate synthase-catalysed reaction and interaction of the enzyme with 2- and-or 4-substituted analogues of dUMP and 5-fluoro-dUMP

Themidylate synthase activity in the development of Hymenolepis diminuta

Phosphorylation of basic amino acid residues in proteins: important but easily missed

Porownanie wlasciwosci syntazy tymidylanowej oczyszczonej z tasiemca szczurzego, Hymenolepis diminuta, z wlasciwosciami enzymu zywiciela wyizolowanego z regenerujacej watraby szczura

Altered mouse leukemia L1210 thymidylate synthase, associated with cell resistance to 5-fluoro-dUrd, is not mutated but rather reflects posttranslational modification

Thymidylate biosynthesis as a target in chemotherapy. Potential importance of a peculiar expression pattern of enzymes involved in thymidylate biosynthesis in nematodes

Comparison of some diaminopyrimidines and amethopterin as inhibitors of insect cell proliferation and of insect folate-metabolizing enzymes

The effect of Arg209 to Lys mutation in mouse thymidylate synthase

Sulfamide antifolates inhibiting thymidylate synthase: synthesis, enzyme inhibition and cytotoxicity

Acyclic analogues of 5-fluoro-dUMP and 5-fluoro-2'-deoxyuridine: Synthesis and inhibition of thymidylate synthase and tumour cell growth

Synthesis and interaction with thymidylate synthase of 5'-dithiophosphate and 5'-fluorothiophosphate of thymidine

Synthesis and biological activity of Na-[4-[N-[[3,4-dihydro-2-methyl-4-oxo-6-quinazolinyl]methyl]-N-propargylamino]phenylacetyl]-L-glutamic acid