title: Expression in E. coli of the catalytic domain of rat poly(ADP-ribose)polymerase.
abstract: 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 alpha-chymotrypsin 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.
authors.affiliation: Centre Hospitalier de l'Université Laval, Laboratoire du métabolisme du poly (ADP-ribose), Endocrinologie Moléculaire, Laurier, Québec, Canada.
authors.forename: J
authors.initials: J
authors.lastname: Thibodeau
authors.forename: F
authors.initials: F
authors.lastname: Simonin
authors.forename: M
authors.initials: M
authors.lastname: Favazza
authors.forename: G
authors.initials: G
authors.lastname: Gradwohl
authors.forename: G
authors.initials: G
authors.lastname: Poirier
authors.forename: G
authors.initials: G
authors.lastname: de Murcia
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D008969:Molecular Sequence Data; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics* / Q000302:isolation & purification; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification; D012689:Sequence Homology, Nucleic Acid
chemical_list: D011994:Recombinant Proteins; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases