Expression in E.coli of the catalytic domain of rat poly(ADP‐ribose)polymerase

FEBS Letters

Gradwohl, Gérard²  Poirier, Guy¹  Simonin, Frédéric²  Favazza, Marisa²  Thibodeau, Jacques¹  de Murcia, Gilbert² 

[1] Centre Hospitalier de l'Université Laval, Laboratoire du métabolisme du poly(ADP-ribose), Endocrinologie Moléculaire, 2705 bout. Laurier, Québec, GIV 4G2, Canada;Institut de Biologie Moléculaire et Cellulaire du CNRS and Centre de Recherche de l'Université Louis Pasteur, 15 rue Descartes, 67084 Strasbourg, France

关键词: Poly(ADP-ribose)polymerase;    E. coli expression;    αChymotrypsin;    NAD+ metabolism;    Immunoblotting;    PARP;    poly(ADP-ribose)polymerase;    kbp;    kilobase pair(s);    PAGE;    polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(90)80770-J

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A 2 kilobase pair cDNA coding for the entire C-terminal catalytic domain of rat poly(ADP-ribose)polymerase has been expressed in E. coli. The overproduced 55 kDa polypeptide is active in synthesizing poly(ADP-ribose) and the 4 kDa N-terminal region of this domain is recognized by the monoclonal antibody C I,2 directed against the calf enzyme. Also, the minor αchymotrypsin cleavage site found in the human catalytic domain is not present in the rat enzyme as revealed by the absence of the 40 kDa specific degradation product in the E. coli cells expressing the rat domain. The expression of this partial rat cDNA should thus permit the rapid purification and subsequent crystallization of the catalytic domain of the enzyme.

【 授权许可】

Unknown