Takashi Shimokawa, Hideki Ogino, Daisuke Maeda, Hitoshi Nakagama, Takashi Sugimura and Mitsuko Masutani

Biochemistry Division, National Cancer Center Research Institute, 5-1-1, Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.

Abstract

Poly(ADP-ribose) polymerase (PARP) polyADP-ribosylates proteins involved in various physiological processes. Accumulated evidence suggests not only protein-conjugated poly(ADP-ribose) but also protein-free poly(ADP-ribose) function in various physiological processes. There are increasing occasions that require protein-free poly(ADP-ribose) to study the function and dynamics of poly(ADP-ribose) in cells. However, the availability of poly(ADP-ribose) is still limited because a chemical synthesis method has not been established. Here, we describe an improved method for the preparation of protein-free poly(ADP-ribose), synthesized enzymatically by using a recombinant PARP-1 expression system and purified with an anion-exchange column chromatography. This method will be useful for biochemical and biological investigation of poly(ADP-ribose) functions and dynamics.