Crystallization and X-ray crystallographic analysis of recombinant chicken poly(ADP-ribose) polymerase catalytic domain produced in Sf9 insect cells

• Published in: Journal of molecular biology Vol. 244; no. 1; pp. 114 - 116
• Main Authors: Jung, S, Miranda, E A, de Murcia, J M, Niedergang, C, Delarue, M, Schulz, G E, de Murcia, G M
• Format: Journal Article
• Language: English
• Published: England 18.11.1994
• Subjects: Animals; Baculoviridae - genetics; Base Sequence; Binding Sites; Chickens; Chromatography, Affinity; Crystallography, X-Ray; Molecular Sequence Data; Peptide Fragments - chemistry; Poly(ADP-ribose) Polymerases - chemistry; Poly(ADP-ribose) Polymerases - genetics; Poly(ADP-ribose) Polymerases - isolation & purification; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Spodoptera - cytology
• ISSN: 0022-2836
• DOI: 10.1006/jmbi.1994.1709

Poly (ADP-ribose) polymerase (PARP) participates in the immediate response in mammalian cells exposed to DNA-damaging agents. Recombinant baculovirus harboring the cDNA of the chicken PARP catalytic domain (40 kDa) have been used to infect Spodoptera frugiperda (Sf9) insect cells. The recombinant polypeptide (30 mg per 1 x 10(9) cells) was purified to homogeneity by 3-aminobenzamide affinity chromatography. The enzymatic properties of the recombinant domain were similar to those of the native fragment. Crystals of the purified recombinant catalytic domain were grown by vapor diffusion. The crystals belong to space group P2(1)2(1)2(1) with unit cell dimensions of a = 59.2 A, b = 65.0 A, c = 96.9 A. They are suitable for X-ray analysis and diffract to 2.0 A.