Expression of human poly(ADP-ribose) polymerase with DNA-dependent enzymatic activity in Escherichia coli

M Ikejima; S Noguchi; R Yamashita; H Suzuki; T Sugimura; M Miwa

doi:10.1016/0006-291x(89)92285-7

Expression of human poly(ADP-ribose) polymerase with DNA-dependent enzymatic activity in Escherichia coli

Biochem Biophys Res Commun. 1989 Sep 15;163(2):739-45. doi: 10.1016/0006-291x(89)92285-7.

Authors

M Ikejima¹, S Noguchi, R Yamashita, H Suzuki, T Sugimura, M Miwa

Affiliation

¹ Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan.

PMID: 2506854
DOI: 10.1016/0006-291x(89)92285-7

Abstract

A cDNA for human poly(ADP-ribose) polymerase was inserted into a plasmid, transfected and expressed in E. coli. A lysate of the E. coli cells containing the expression plasmid reacted with antibody against human poly(ADP-ribose) polymerase and synthesized poly(ADP-ribose). The partially purified poly(ADP-ribose) polymerase expressed in E. coli had the same molecular weight and enzymological properties as human placental poly(ADP-ribose) polymerase, including affinity for NAD, turnover number and DNA-dependency for activity. This expression system should be useful for structure-function analysis of poly(ADP-ribose) polymerase.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Blotting, Western
DNA / genetics
Electrophoresis, Polyacrylamide Gel
Escherichia coli / enzymology*
Gene Expression Regulation*
Humans
NAD / metabolism
Poly(ADP-ribose) Polymerases / genetics*
Transfection

Substances

NAD
DNA
Poly(ADP-ribose) Polymerases