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

FEBS Lett. 1990 May 7;264(1):81-3. doi: 10.1016/0014-5793(90)80770-j.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cloning, Molecular
  • DNA / genetics
  • Escherichia coli / enzymology
  • Escherichia coli / genetics*
  • Gene Expression
  • Genes
  • Molecular Sequence Data
  • Poly(ADP-ribose) Polymerases / genetics*
  • Poly(ADP-ribose) Polymerases / isolation & purification
  • Rats
  • Recombinant Proteins / isolation & purification
  • Sequence Homology, Nucleic Acid

Substances

  • Recombinant Proteins
  • DNA
  • Poly(ADP-ribose) Polymerases