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Antimicrobial Agents and Chemotherapy, June 2002, p. 2035-2037, Vol. 46, No. 6
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.6.2035-2037.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Nonradiolabeling Assay for WaaP, an Essential Sugar Kinase Involved in Biosynthesis of Core Lipopolysaccharide of Pseudomonas aeruginosa

Xin Zhao, Cory Q. Wenzel, and Joseph S. Lam*

Canadian Bacterial Diseases Network, Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada

Received 7 December 2001/ Returned for modification 9 January 2002/ Accepted 13 March 2002

waaP is present in the lipopolysaccharide (LPS) core gene clusters of a wide range of gram-negative bacteria, and is an essential gene in Pseudomonas aeruginosa. The WaaP protein is a sugar kinase that adds phosphate to heptose I in the core oligosaccharide. This study describes the standardization and utility of a chemiluminescence-based enzyme-linked immunosorbent assay for the detection of WaaP kinase activity. Important features of the assay include high sensitivity, the preparation of dephosphorylated LPS as a substrate, and the use of monoclonal antibody 7-4 that specifically recognizes phosphate substituents in the LPS core.

* Corresponding author. Mailing address: Canadian Bacterial Diseases Network, Department of Microbiology, University of Guelph, Guelph, ON N1G 2W1, Canada. Phone: (519) 824-4120, ext. 3823. Fax: (519) 837-1802. E-mail: jlam{at}uoguelph.ca.

Antimicrobial Agents and Chemotherapy, June 2002, p. 2035-2037, Vol. 46, No. 6
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.6.2035-2037.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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