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. 1989 Jul;57(7):1873–1878. doi: 10.1128/iai.57.7.1873-1878.1989

Effects of eliminating a disulfide bridge within domain II of Pseudomonas aeruginosa exotoxin A.

I H Madshus 1, R J Collier 1
PMCID: PMC313813  PMID: 2499539

Abstract

Cysteines 265 and 287 of Pseudomonas aeruginosa exotoxin A (ETA) were substituted by serine, thereby eliminating a disulfide bridge within domain II, the putative membrane insertion-translocation domain. Purified mutant toxin was 80-fold less toxic for mouse L cells than was wild-type ETA while retaining the same specific activity in the ADP-ribosyltransferase reaction as did wild-type toxin. Binding of the nonionic detergent Triton X-114 by mutant ETA occurred at a slightly higher pH than did binding by wild-type ETA, suggesting that the mutant protein more readily undergoes a conformational change exposing hydrophobic regions. Data are presented supporting the notion that the mutant and wild-type toxins enter from the same intracellular compartment. The lower cytotoxicity of the mutant protein could be due to accelerated intracellular degradation or abortive, premature membrane insertion.

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Selected References

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