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Expression in E.coli of the catalytic domain of rat poly(ADP‐ribose)polymerase
Author(s) -
Thibodeau Jacques,
Simonin Frédéric,
Favazza Marisa,
Gradwohl Gérard,
Poirier Guy,
de Murcia Gilbert
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80770-j
Subject(s) - microbiology and biotechnology , enzyme , complementary dna , polymerase , poly adp ribose polymerase , biochemistry , biology , chemistry , gene
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.