New adsorbents for thymidylate synthase affinity chromatography
New affinity adsorbents, intended for chromatography of thymidylate synthase (EC 2.1.1.45) from different sources, consisting of p-[N-[(2-amino-4-hydroxy-6-quinazolinyl)-methyl]-N-2-propynylamino]benzoyl-γ-[α-(3-carboxypropylamino)]glutamyl-glutamyl immobilized either on macroporous copolymer of acrylonitrile and n-butyl acrylate or on macroporous polymer of acrylonitrile itself, both crosslinked with divinylbenzene and having aminoethyl groups, were obtained. Both adsorbents were found to be effective in dUMP-dependent binding of thymidylate synthase from regenerating rat liver.
Thymidylate synthases from Hymenolepis diminuta and regenerating rat liver: purification, properties, and inhibition by substrate and cofactor analogues.
Comparative studies of thymidylate synthases, isolated from the tapeworm, Hymenolepis diminuta, and regenerating liver of its host, rat, aimed at a possibility of specific inhibition of the helminthic enzyme, are presented. While similar in structure (dimers with monomer molecular masses of 33.7 kDa and 34.9 kDa, respectively) and parameters describing interactions with substrates and products, the tapeworm and rat enzymes differed in the dependences of reaction velocity on temperature (Arrhenius plots biphasic and linear, respectively). The tapeworm, compared with the host, enzyme was less sensitive to the competitive slow-binding inhibition by 5-fluoro-dUMP and its 2-thio congener, but eq…