Investigation into the catalytic role for the tryptophan residues within domain III of Pseudomonas aeruginosa exotoxin A

Biochemistry
B K BeattieA Rod Merrill

Abstract

The role of the tryptophan residues in the substrate-binding and catalytic mechanism of an enzymatically active C-terminal fragment of Pseudomonas aeruginosa exotoxin A was studied by individually or jointly replacing these residues with phenylalanine. Substitution of W-466 decreased the ADP-ribosyltransferase and NAD(+)-glycohydrolase activities by 20- and 3-fold, respectively. In contrast, substitution of W-417 or W-558 with phenylalanine both resulted in a 3-fold decrease in ADP-ribosyltransferase activity with, however, only a decrease by 40% and 70% in NAD(+)-glycohydrolase activity, respectively. Simultaneous replacement of W-466 and W-558 resulted in a 200-fold decrease in ADP-ribosyltransferase and an 6-fold decrease in NAD(+)-glycohydrolase activities, suggesting that W-466 may play a minor role in the transfer of ADP-ribose to the eEF-2 protein. Chemical modification of the tryptophan residues in the wild-type toxin fragment by N-bromosuccinimide revealed the presence of a single residue important for enzymatic activity, W-466, with a minor contribution from W-558. Additionally, tryptophan residues, W-305 and W-417, were refractory to oxidation by N-bromosuccinimide, which likely indicated the buried nature of these r...Continue Reading

References

Mar 28, 1977·Biochimica Et Biophysica Acta·A Michel, J Dirkx
May 1, 1986·The Journal of Pediatrics·M L Vasil

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Citations

Apr 27, 2012·The Journal of Biological Chemistry·Robert J FieldhouseA Rod Merrill
Sep 13, 2012·The Journal of Biological Chemistry·Adin ShnifferA Rod Merrill
Dec 21, 2010·PLoS Computational Biology·Robert J FieldhouseA Rod Merrill
Oct 25, 2008·International Journal of Medical Microbiology : IJMM·Philipp Wolf, Ursula Elsässer-Beile
Jan 13, 2006·Trends in Biochemical Sciences·Susan P YatesA Rod Merrill
Oct 7, 2015·Frontiers in Microbiology·Marta Michalska, Philipp Wolf
Apr 2, 2015·Biochimie·Vicky M-H Sung
May 17, 2006·Technology in Cancer Research & Treatment·Kenji MuroJeffrey J Raizer
Aug 19, 2005·Nature·René JørgensenGregers R Andersen
Aug 18, 2005·The Journal of Biological Chemistry·Brooke D LazarusJohn A Hanover
Jul 18, 2001·The Journal of Biological Chemistry·S P Yates, A R Merrill
May 21, 1999·The Journal of Biological Chemistry·B K Beattie, A R Merrill
Aug 19, 2004·The Journal of Biological Chemistry·René JørgensenGregers Rom Andersen
Mar 16, 2017·JCI Insight·Christoph T Ellebrecht, Aimee S Payne

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