Benzamide affinity matrix

Fig. 1. The binding of coenzymic DNA to benzamide affinity matrix. Ordinate auto-poly(ADP-ribosylation) of the enzyme abscissa time of enzymic reaction. -- Recovery of free coen-...

Isolation of calf thymus poly(ADP-ribose) polymerase (at least 95% homogeneous, specific activity = 1500 nmol ADP-ribose incorporated mg protein in 1 min) and coenzymic DNA were carried out using published methods [8]. The synthesis of 0-amino-benzamide agarose affinity matrix, DNA-cellulose, the method of iodination of the enzyme protein (with the preparation of the Bolton-Hunter reagent adduct of... 

Benzamide covalently bound to agarose matrix (linker at the ortoposition) does not bind poly(ADP-ribose) polymerase (data not shown). However, the binding to this affinity matrix of the coenzymic DNA of poly(ADP-ribose) polymerase is readily demonstrable (Fig. 1). The free coenzymic DNA was determined by poly(ADP-ribose) polymerase, an experiment based on the linear rate of enzyme-autopoly(ADP-ribosyla-tion) that is directly proportional to the concentration of free coenzymic DNA in the... 

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