Expression of human poly(ADP-ribose) polymerase with DNA-dependent enzymatic activity in Escherichia coli.
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.
Ikejima
M
M
Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan.
Noguchi
S
S
Yamashita
R
R
Suzuki
H
H
Sugimura
T
T
Miwa
M
M
D015153:Blotting, Western; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D005786:Gene Expression Regulation; D006801:Humans; D009243:NAD / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics*; D014162:Transfection
D009243:NAD; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases