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. 1992 Mar;174(6):2032–2038. doi: 10.1128/jb.174.6.2032-2038.1992

Localization of diphtheria toxin nuclease activity to fragment A.

S L Lessnick 1, J B Lyczak 1, C Bruce 1, D G Lewis 1, P S Kim 1, M L Stolowitz 1, L Hood 1, B J Wisnieski 1
PMCID: PMC205811  PMID: 1548241

Abstract

We describe a series of experiments that aimed to establish whether nuclease activity is actually associated with diphtheria toxin (DTx) and its A subunit (DTA), as we originally reported (M. P. Chang, R. L. Baldwin, C. Bruce, and B. J. Wisnieski, Science 246:1165-1168, 1989). Here we show that (i) trypsinization of DTx does indeed produce nucleolytically active DTA, (ii) reduction of electroeluted, unreduced, cleaved DTx (58 kDa) yields nuclease-active DTA (24 kDa), and (iii) fractionation of DTx and DTA by anion-exchange chromatography leads to coelution of nuclease activity with both forms of the toxin, even though each form elutes at a distinct salt concentration. In addition, we show that Escherichia coli-derived DTA also expresses nuclease activity. These studies confirm our initial assertion that the nuclease activity observed in DTx preparations is intrinsic to the DTA portion of DTx.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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