期刊
ANALYTICAL BIOCHEMISTRY
卷 353, 期 2, 页码 266-271出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2006.03.043
关键词
toxin; nonradioactive; inhibition of protein synthesis; pseudomonas; ADP-ribosylation; reticulocyte
资金
- Intramural NIH HHS Funding Source: Medline
Pseudomonas exotoxin A (PE) inhibits protein synthesis by NAD-dependent ADP-ribosylation of eukaryotic elongation factor 2. Traditionally, toxin activity has been characterized, either in living cells or cell-free systems, using radioactive, compounds for quantification. The increased costs of radioactive waste disposal together with heightened security concerns have made the use of radioactive isotopes less attractive for routine laboratory assays. We therefore adapted a cell-free rabbit reticulocyte in vitro transcription-translation system that utilizes a reporter (beta-galactosidase) to measure toxin activity. The assay for PE is rapid, scalable, log-linear, NAD dependent and can be used to assess the neutralizing activity of anti-PE antibody preparations. (c) 2006 Elsevier Inc. All rights reserved.
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