Rapid assay of poly(ADP-ribose) glycohydrolase.
We have developed a rapid, highly reproducible assay to determine poly(ADP-ribose) glycohydrolase activity which measures directly the appearance of the reaction product. We also analysed the majority of different techniques which are used to determine poly(ADP-ribose) glycohydrolase activity and found that the apparent activity can vary extensively depending on the method used. Thin-layer chromatography using PEI-F-cellulose was the only method which evaluated directly the specific release of ADP-ribose; by comparison with this method, the other procedures gave an over- or under-estimation of 2- to 10-fold of the enzymatic activity. A rapid method of affinity chromatography has also been developed to synthesize and purify in high yield poly(ADP-ribose) (35% conversion of 1 mM NAD to poly(ADP-ribose)).
Ménard
L
L
Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Qué., Canada.
Poirier
G G
GG
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose / Q000096:biosynthesis / Q000302:isolation & purification; D011865:Radioisotope Dilution Technique; D013950:Thymus Gland / Q000201:enzymology*
D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose; D006026:Glycoside Hydrolases; C022407:poly ADP-ribose glycohydrolase