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- W2063742699 abstract "Poly (A) synthetase from mouse ascites cells. I. Purification and characterization of the enzyme 1. The poly(A) synthetase from mouse ascites cells which incorporates [14C]AMP into a preexisting polyribonucleotide chain is a weakly acidic protein (isoelectric point = 6 ± 0.05) with a molecular weight, determined by sucrose gradient centrifugation, close to 80 000. Its exclusion from Sephadex G-200 would indicate that it can exist in a complex form. Incubation of the enzyme in a medium of high ionic strength (0.5 M KCl) results in a reversible dissociation. However, centrifugation in a high gravitational field (284 000 × g) in presence of 0.5 M KCl leads to an irreversible dissociation of the enzyme. 2. The enzyme is heat labile and loses 50 % of its activity in 4 min at 45 °C. The polyribonucleic acid primer and ATP have distinct roles in the stabilisation of the enzyme. 3. The enzyme was 150-fold purified. Nevertheless, it still contains trace amounts of nucleic acids and has slight nuclease activity. Purification of the enzyme results in the appearance of new properties, which are discussed in the second paper. 1. La poly(A) synthétase des cellules d'ascite de souris, responsable de l'incorporation de [14C]AMP sur une chaîne polyribonucléotidique préexistante, est une protéine faiblement acide (point isoélectrique = 6 ± 0.05), de pois moléculaire voisin de 80 000. Mais son exclusion d'une colonne de Sephadex G-200 notamment, indiquerait qu'elle peut exister sous forme plus complexe. La vitesse de sédimentation de la protéine enzymatique est abaissée à force ionique élevée. Cette dissociation peut être réversible. Mais la composition en sous-unités ne peut être actuellement affirmée. 2. La protéine enzymatique est thermosensible: elle perd 50 % de son activité après chauffage pendant 4 min à 45 °C. L'ATP et l'amorceur polyribonucléotidique ont des rôles distincts dans la stabilisation de la structure de la protéine. 3. La protéine enzymatique a été purifiée environ 150 fois mais la fraction purifiée contient encore des traces d'acides nucléiques et d'activité nucléasique. La purification a entraîné des modifications de l'activité enzymatique et l'apparition de propriétés nouvelles dont l'étude fait l'objet de l'article suivant." @default.
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- W2063742699 date "1972-12-01" @default.
- W2063742699 modified "2023-10-17" @default.
- W2063742699 title "Polyadénylate synthétase des cellules d'ascite de souris" @default.
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- W2063742699 doi "https://doi.org/10.1016/0005-2787(72)90287-0" @default.
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