Expression in yeast and purification of functional recombinant human poly(ADP-ribose)polymerase (PARP). Comparative pharmacological profile with that of the rat enzyme

J Enzyme Inhib. 2000;15(5):461-9. doi: 10.3109/14756360009040702.

Abstract

Human poly(ADP-ribose)polymerase (PARP) was expressed in the yeast line JEL1 under the control of a GAL promoter. Proteins were extracted and human recombinant PARP purified to apparent homogeneity. The pharmacological profile of this human enzyme was characterised in terms of the effects of known inhibitors of PARP belonging to various chemical families and this was compared with that of the rat enzyme purified from rat testes, using the same purification protocol. The rat and the human enzymes appeared very similar in terms of their sensitivities to those selected inhibitors.

Publication types

  • Comparative Study

MeSH terms

  • 1-Naphthylamine / analogs & derivatives*
  • 1-Naphthylamine / pharmacology
  • Animals
  • Benzamides / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors / pharmacology*
  • Gene Expression
  • Humans
  • Kinetics
  • Male
  • Naphthalimides
  • Phenanthrenes / pharmacology
  • Poly(ADP-ribose) Polymerase Inhibitors*
  • Poly(ADP-ribose) Polymerases / genetics
  • Poly(ADP-ribose) Polymerases / isolation & purification
  • Quinazolines / pharmacology
  • Quinolones / pharmacology
  • Rats
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Saccharomyces / enzymology*
  • Sensitivity and Specificity
  • Testis / enzymology*

Substances

  • Benzamides
  • Enzyme Inhibitors
  • Naphthalimides
  • Phenanthrenes
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Quinazolines
  • Quinolones
  • Recombinant Proteins
  • 4-amino-1,8-naphthalimide
  • 1-Naphthylamine
  • phenanthridone
  • Poly(ADP-ribose) Polymerases