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. 1990 Aug 25;18(16):4691–4694. doi: 10.1093/nar/18.16.4691

An affinity matrix for the purification of poly(ADP-ribose) glycohydrolase.

H Thomassin 1, M K Jacobson 1, J Guay 1, A Verreault 1, N Aboul-ela 1, L Menard 1, G G Poirier 1
PMCID: PMC331920  PMID: 2395636

Abstract

The preparation of quantities of poly(ADP-ribose) glycohydrolase sufficient for detailed structural and enzymatic characterizations has been difficult due to the very low tissue content of the enzyme and its lability in late stages of purification. To date, the only purification of this enzyme to apparent homogeneity has involved a procedure requiring 6 column chromatographic steps. Described here is the preparation of an affinity matrix which consists of ADP-ribose polymers bound to dihydroxyboronyl sepharose. An application is described for the purification of poly(ADP-ribose) glycohydrolase from calf thymus in which a single rapid affinity step was used to replace 3 column chromatographic steps yielding enzyme of greater than 90% purity with a 3 fold increase in yield. This matrix should also prove useful for other studies of ADP-ribose polymer metabolism and related clinical conditions.

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Selected References

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