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1.
Bioorg Khim ; 25(4): 282-9, 1999 Apr.
Artigo em Russo | MEDLINE | ID: mdl-10422595

RESUMO

The effect of core trimers, (2'-5')-analogues of oligoadenylic acid containing 9-(3-deoxy-3-fluoro-beta-D-xylofuranosyl)adenine (AF) and 3'-deoxy-3'-fluoroadenosine (AF) in various positions of the oligomer chain, on the lytic activity of human natural killer cells (NK cells) was studied in three different ways. The cellular cytotoxicity was determined using a highly sensitive nonradioactive approach employing a chelate europium-diethylenetriamino-pentaacetic acid complex (Eu-DTPA). It was shown that all fluorodeoxyanalogues enhance the lytic activity of intact NK lymphocytes, which follows from the lysis rate constant k2. At the same time, the substitution of either the central adenosine fragment or (to a greater extent) the 5'-terminal residue of (2'-5')A3 with AF causes a decrease in the number of active NK cells, which, unlike the case of the natural core trimer, leads to a loss of the capacity to increase the activity of NK. By contrast, isomeric ribo-analogues. (2'-5')(AF)A2 and (2'-5')A(AF)A, and trimers with the 2'(3')-terminal nucleotide substituted by AF or AF increased the activity of NK cells with an effectiveness close to or higher than the natural trimer (2'-5')A3. Inasmuch as isomeric xylo- and ribo-3'-deoxy-3'-fluoroanalogues of (2'-5')A3 are stereochemically modified oligomers, the data unambiguously suggest that the spatial structure of these trimers affects the increase in the lytic activity of NK cells.


Assuntos
Citotoxicidade Imunológica/efeitos dos fármacos , Desoxiadenosinas/farmacologia , Células Matadoras Naturais/imunologia , Humanos , Células Matadoras Naturais/química
2.
Bioorg Khim ; 25(10): 763-7, 1999 Oct.
Artigo em Russo | MEDLINE | ID: mdl-10645479

RESUMO

Mouse antibodies to (2'-5')oligoadenylates were obtained by the immunization of animals with the (2'-5')oligoadenylic acid trimer conjugated with bovine serum albumin through a 2',3'-levulinic acid residue. Using radioimmunoassay, the reactivity of mouse polyclonal antibodies to the (2'-5')oligoadenylic acid trimer was studied for the trimer analogues containing 9-(3-deoxy-3-fluro-beta-D- xylofuranosyl)adenine and 3'-deoxy-3'-fluoro-adenosine in various positions of the chain. It was found that (a) the three-dimensional structure of short oligonucleotides is an important factor in the antibody recognition; (b) antibodies are more sensitive to modifications of the 5'-terminal and central ribose fragments of the (2'-5')oligoadenylic acid trimer; (c) the 3'-hydroxyl group plays a secondary role in the formation of the antigen determinant.


Assuntos
Nucleotídeos de Adenina/imunologia , Anticorpos/imunologia , Oligorribonucleotídeos/imunologia , Nucleotídeos de Adenina/química , Animais , Anticorpos/química , Biopolímeros , Dicroísmo Circular , Camundongos , Oligorribonucleotídeos/química , Soroalbumina Bovina/imunologia , Estereoisomerismo
3.
Acta Biochim Pol ; 45(1): 87-94, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9701500

RESUMO

Two non-conventional analogues of ATP, 3'-deoxyadenosine-2'-triphosphate (3'-d-2'-ATP) and 2'-deoxyadenosine-3'-triphosphate (2'-d-3'-ATP), the syntheses of which are described, were examined as potential phosphate donors for the nucleoside kinases: 2'-deoxycytidine kinase (dCK), cytosolic thymidine kinase (TK1) and mitochondrial thymidine kinase (TK2). The reactions were monitored by means of a mixture of [gamma-32P]ATP and cold analogue, and/or with the use of 3H-labelled acceptors and cold donor. With dCK, using equimolar mixtures of ATP with each analogue, and dC as acceptor, phosphate transfer from 3'-d-2'-ATP and 2'-d-3'-ATP amounted to 34% and 14%, respectively. With each analogue used alone (each at concentration of 100 microM), phosphate transfer from 3'-d-2'-ATP was 55% that from ATP, and from 2'-d-3'-ATP 16%. With human TK2, and equimolar mixtures of [gamma-32P]ATP with each of the analogues, and 1 microM dT as acceptor, there was no detectable transfer from either analogue. But, when each analogue was used alone, phosphate transfer attained 11% and 5%, respectively, that for ATP alone. With the low affinity form of human TK1, and dT as acceptor, only low phosphate transfer occurred with either analogue used alone. Both compounds exhibited Michaelis-Menten kinetics (with significantly lower Vmax than ATP), while ATP exhibited cooperative kinetics with all three kinases.


Assuntos
Nucleotídeos de Desoxiadenina/química , Desoxicitidina Quinase/química , Timidina Quinase/química , Catálise , Humanos , Fosforilação
4.
Biochem Biophys Res Commun ; 245(2): 430-4, 1998 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-9571169

RESUMO

To elucidate further the roles played by the adenine bases in the interaction of RNase L (EC 3.1.2.6) with the 2',5'-oligoadenylate 2-5A, p5'A2'(p5'A2')np5' A, a series of sequence-specific 1-deazaadenosine (c1A)-substituted analogues were synthesized and evaluated for their ability to bind to and activate human RNase L in comparison to earlier reported inosine-substituted congeners of 2-5A. Substitution of only the 5'-terminal adenosine of p5'A2'p5'A2 p5 A with c1A afforded an analogue with strongly diminished RNase L binding and activation ability, while replacement of the second or middle adenosine of p5 A2' p5'A2'p5' A had only a modest effect. In distinct contrast to p5'A2'p5'A2'p5'I, the c1A analogue with the third or 2'-terminal adenosine replacement approached parent p5' A2'p5'A2'p5' A in RNase L activation ability. These results permitted a further dissection of the role of various nucleotidic functional groups in the interaction of 2-5A with RNase L: specifically, that the 5'-terminal adenosine purine N-1 moiety is key for binding to RNase L, while the 2'-terminal adenosine N-6 exocyclic amino group is critical for RNase L activation.


Assuntos
Nucleotídeos de Adenina/farmacologia , Endorribonucleases/metabolismo , Oligorribonucleotídeos/farmacologia , Adenosina/análogos & derivados , Adenosina/metabolismo , Ativação Enzimática/fisiologia , Humanos , Estrutura Molecular , Oligorribonucleotídeos/metabolismo , Proteínas de Ligação a RNA , Proteínas Recombinantes/metabolismo , Tubercidina/química
6.
Eur J Biochem ; 221(2): 759-68, 1994 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-8174555

RESUMO

A one- and two-dimensional NMR study has been performed on seven A(2'-5')A(2'-5')A fragments containing 9-(3'-fluoro-3'-deoxy-beta-D-xylofuranosyl)-adenine (AF) or 3'-fluoro-3'-deoxyadenosine (AF) residues at different positions, and on the corresponding monomers. A(2'-5')A(2'-5')A served as a reference compound. The fluoro substituent governs the conformation of the sugar ring: an AF residue displays mainly N-type sugar and the ring is considerably flattened (phi N approximately 30 degrees) compared to AF residues (phi S approximately 40 degrees), which exhibit almost pure S-type conformation. Moreover, in AF moieties the rotamer distribution around torsion angle gamma (O5'-C5'-C4'-C3') and the base orientation are influenced to a large extent by the presence of the fluorine substituent. The sugar rings of nonfluorinated residues in the trimers appear rather flexible. A possible correlation between the conformational characteristics of the fluorinated fragments and their biological activity has been found: the fragments that meet the prerequisites for binding to RNase L indeed show enhanced binding to this endonuclease. Furthermore, substitution of the 3'-OH group of the second residue by hydrogen or of the 3'-OH group of the 2'-terminal residue by fluorine or hydrogen results in increased resistance towards 2'-5'-phosphodiesterase.


Assuntos
Nucleotídeos de Adenina/química , Desoxiadenosinas/química , Endorribonucleases/metabolismo , Nucleotídeos de Adenina/farmacologia , Desoxiadenosinas/farmacologia , Espectroscopia de Ressonância Magnética , Conformação Molecular , Polímeros , Relação Estrutura-Atividade
7.
Biochem Biophys Res Commun ; 167(1): 20-6, 1990 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-2155612

RESUMO

Analogs of 2-5A trimer 5'-monophosphate (2'-5')pA3,p5'A2'p5'A2'p5'A containing 9-(3-fluoro-3-deoxy-c-D-xylofuranosyl)adenine (AF) or 3'-fluoro-3'- deoxyadenosine (AF) at different positions of the chain have been synthesized. All of them were compared with (2'-5')pA3 and (2'-5')pA2 (3'dA) by (i) their ability to bind to 2-5A-dependent endoribonuclease(RNase L) of mouse L cells and of rabbit reticulocyte lysates and (ii) their susceptibility to the degradation by the (2'-5')phosphodiesterase activity. The results of this study suggest that the oligonucleotide conformation is important for its biochemical properties.


Assuntos
Nucleotídeos de Adenina/metabolismo , Endorribonucleases/metabolismo , Oligorribonucleotídeos/metabolismo , Diester Fosfórico Hidrolases/metabolismo , Animais , Células Cultivadas , Camundongos , Coelhos
8.
Bioorg Khim ; 14(9): 1157-61, 1988 Sep.
Artigo em Russo | MEDLINE | ID: mdl-3219131

RESUMO

The substrate specifity of adenosine deaminase has been studied using C'-methyl derivatives of adenosine. On the basis of the correlation revealed between conformations of 2'- and 3'-C-methyladenosine and their substrate properties, a modified stereochemical model is suggested: the enzyme accepts the substrate within a N-type conformational range (4E----4T3----3E) of the furanose ring. The model was analysed in details using a number of C3'-modified adenosines and 5'-C-methyladenosine analogues with D-allo- and L-talo-configuration.


Assuntos
Adenosina Desaminase/metabolismo , Adenosina/metabolismo , Nucleosídeo Desaminases/metabolismo , Adenosina/análogos & derivados , Fenômenos Químicos , Química , Cinética , Especificidade por Substrato
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