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

• Autorzy: Ciesla J , Machnicka B. , Pawelczak K. , Rzeszotarska B. , Rode W.
• Języki publikacji: PL

Abstrakty

EN

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.

Słowa kluczowe

PL

szczury; pasozyty zwierzat; syntaza tymidylanowa; parazytologia; enzymy; watroba; zywiciele; Hymenolepis diminuta; tasiemce

• Wydawca: -
• Czasopismo: Wiadomości Parazytologiczne
• Rocznik: 1992
• Tom: 38
• Numer: 1-2
• Opis fizyczny: s.23-29, rys.,tab.,bibliogr.

Twórcy

autor

Ciesla J

• Instytut Biologii Doswiadczalnej PAN, ul.Pasteura 3, 02-093 Warszawa

autor

Machnicka B.

autor

Pawelczak K.

autor

Rzeszotarska B.

autor

Rode W.

Bibliografia

• CIEŚLA J., ZIELIŃSKI Z., MACHNICKA B., RODE W. 1987. Thymidylate synthase activity in the development of Hymenolepis diminuta. Acta Biochim. Polon. 34: 291-298.
• DANENBERG F. V. 1977. Thymidylate synthase - a target enzyme in cancer chemotherapy. Biochim. Biophys. Acta 473: 73-92.
• HEIDELBERGER C., DANENBERG P. V., MORAN R. G. 1983. Fluorinated pyrimidines and their nucleosides. In: A. MEISTER [Ed.] Advances in Enzymology, JOHN WILEY & Sons, New York, 54: 57-119.
• HIGGINS G. M., ANDERSON R. M. 1931. Experimental pathology of the liver. Arch. Pathol. 12: 186-202.
• JASTREBOFF M., KĘDZIERSKA B., RODE W. 1982. Properties of thymidylate synthetase from EHRLICH ascites carcinoma cells. Biochem. Pharmacol. 31: 217-223.
• RODE W., KULIKOWSKI T., KĘDZIERSKA B., SHUGAR D. 1987. Studies on the interaction with thymidylate synthase of analogues of 2' -deoxyuridine-5' -phosphate and 5-fluoro-2' -deoxyuridine-5' -phosphate with modilied phosphate groups. Ibid. 36: 203-210.
• -., SCANLON K. J., HYNES J., BERTINO J. R. 1979. Purification of mammalian tumor (L1210) thymidylate synthetase by affinity chromatography on stable biospecilic adsorbent. J. Biol. Chem. 254: 11538-11543.
• SANTI D. V. 1980. Perspectives on the design and biochemical pharmacology of inhibitors of thymidylate synthetase. J. Med. Chem. 23: 103-111.
• SPECTOR T. 1978. Refinement of the coomasie blue method of protein quantitation. Anat. Biochem. 86: 142-146.