Biomedical subjects

S Izuta

Publications and source records attributed to S Izuta.

40 records · Page 3Linked to original sources

PubMed1986

Recognition of sugar moieties on substrate analogues by DNA polymerase alpha 2-primase from developing cherry salmon (Oncorhynchus masou) testes.

Several dCTP or dATP analogues, bearing an azido or amino group on 2'- or 3'-position of its sugar moiety, were examined for their inhibitory effects on DNA polymerase alpha 2-primase from developing cherry salmon (Oncorhynchus masou) testes, and the recognition of sugar moieties of the analogues by primase and related nucleic acid polymerases were compared. Among the dCTP analogues tested, 2'-azido-2',3'-dideoxy CTP inhibited primase strongly and RNA polymerases I and II to lesser extent. Although, the Ki value for primase was larger than those of RNA polymerases, the Ki/Km value for primase was smaller. In contrast, 3'-amino-2',3'-dideoxy CTP selectively inhibited DNA polymerase beta. In dATP analogue series, 3'-amino-3'-deoxy ATP inhibited RNA polymerases I and II very strongly to the same extent as 3'-deoxy ATP. This analogues was a more selective inhibitor for RNA polymerases I and II than 3'-dATP itself.

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PubMed1985

Inhibitory effects of various 3'-deoxyribonucleotides on DNA polymerase alpha 2-primase from developing cherry salmon (Oncorhynchus masou) testes.

DNA polymerase alpha 2-primase has been purified 2750 fold from developing cherry salmon (Oncorhynchus masou) testes by the following purification steps: fractional extraction, phosphocellulose (1st), ammonium sulfate fractionation, DEAE-cellulose, phosphocellulose (2nd), hydroxylapatite and single-stranded DNA-cellulose column chromatographies. Final preparation of this enzyme has a specific activity of 107,000 units/mg protein (activated salmon sperm DNA as template-primer). DNA primase activity (rGTP dependent incorporation of labelled dGMP into poly (dC) or rNTP dependent incorporation of dNMP into M13 single-stranded DNA) was tightly associated with DNA polymerase alpha activity during all stage of this purification process. Inhibition of DNA primase activity by six kinds of 3'-deoxyribonucleotides was studied by using rNTP dependent DNA synthesis on M13 DNA as template. The inhibition constants (Ki) were larger than those of DNA-dependent RNA polymerases I and II. However, Ki/Km values were very close.

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PubMed1984

Inhibitory effects of 1-beta-D-arabinofuranosyl-5-styryluracil 5'-triphosphates on DNA-dependent DNA polymerases alpha and beta from developing cherry salmon (Oncorhynchus masou) testes: on the approach to the selective affinity chromatography for DNA polymerase alpha.

Starting from 1-beta-D-arabinofuranosyluracil, several 5-substituted derivatives were synthesized via 5-mercuri intermediate. The resulting nucleosides were converted into their corresponding 5'-triphosphates and examined for their inhibitory effects on DNA-dependent DNA polymerases alpha and beta from developing cherry salmon (Oncorhynchus masou) testes. The following results were obtained. All the compounds tested showed remarkable inhibitory effects on DNA polymerase alpha, but lesser extent on DNA polymerase beta. The inhibitory action of the hydrophobic 5-substituents against DNA polymerase alpha was stronger than against DNA polymerase beta. For example, Ki/Ki of 5-(E)-3-amino-styryl-Ara UTP is 0.32 and Ki/Km for pol alpha/Ki/Km for pol beta is 0.19. For that reason, we chose this compound as a candidate for a ligand which is applicable to selective affinity chromatography for DNA polymerase alpha.

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PubMed1983

A novel acid ribonuclease from white-spotted charr (Salvelinus leucomaenis): purification and characterization of the homogeneous enzyme.

An novel acid ribonuclease has been purified approx. 1300 fold from acid extract of developing white-spotted charr (Salvelinus leucomaenis) testes. The final preparation of the enzyme was homogeneous on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS). The molecular weight was determined to be 20,000 with both Sephadex gel filtration and SDS-polyacrylamide gel electrophoresis. The isoelectric point was determined to be 6.4 and the hydrolytic activity was stimulated with low concentration of phosphate. The enzyme has an optimal pH at 5.5 and hydrolyzed yeast RNA and several synthetic homopolyribonucleotides. Under appropriate conditions, the enzyme hydrolyzed substrates, yeast RNA, poly (I), poly (A) and poly (U), in a ratio of 100:900:30:20, respectively. However, poly (C) could not be hydrolyzed under any conditions tested so far. Yeast RNA was hydrolyzed into monoto pentanucleotides. The range of substrate-specificity of this enzyme will be discussed on the basis of the analysis of these products.

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