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Journal of Bacteriology, January 2006, p. 191-201, Vol. 188, No. 1
0021-9193/06/$08.00+0     doi:10.1128/JB.188.1.191-201.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Pseudomonas aeruginosa Lipid A Deacylase: Selection for Expression and Loss within the Cystic Fibrosis Airway

Robert K. Ernst,1* Kristin N. Adams,1 Samuel M. Moskowitz,2 Gretchen M. Kraig,1 Kiyoshi Kawasaki,3,5 Christopher M. Stead,6 M. Stephen Trent,6 and Samuel I. Miller1,3,4

Departments of Medicine,1 Pediatrics,2 Microbiology,3 Genome Sciences, University of Washington, Seattle, Washington 98195,4 Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan,5 Department of Microbiology, J. H. Quillen College of Medicine, Johnson City, Tennessee 376146

Received 13 July 2005/ Accepted 10 October 2005

Lipopolysaccharide (LPS) is the major surface component of gram-negative bacteria, and a component of LPS, lipid A, is recognized by the innate immune system through the Toll-like receptor 4/MD-2 complex. Pseudomonas aeruginosa, an environmental gram-negative bacterium that opportunistically infects the respiratory tracts of patients with cystic fibrosis (CF), can synthesize various structures of lipid A. Lipid A from P. aeruginosa strains isolated from infants with CF has a specific structure that includes the removal of the 3 position 3-OH C10 fatty acid. Here we demonstrate increased expression of the P. aeruginosa lipid A 3-O-deacylase (PagL) in isolates from CF infants compared to that in environmental isolates. PagL activity was increased in environmental isolates by growth in medium limited for magnesium and decreased by growth at low temperature in laboratory-adapted strains of P. aeruginosa. P. aeruginosa PagL was shown to be an outer membrane protein by isopycnic density gradient centrifugation. Heterologous expression of P. aeruginosa pagL in Salmonella enterica serovar Typhimurium and Escherichia coli resulted in removal of the 3-OH C14 fatty acid from lipid A, indicating that P. aeruginosa PagL recognizes either 3-OH C10 or 3-OH C14. Finally, deacylated lipid A species were not observed in some clinical P. aeruginosa isolates from patients with severe pulmonary disease, suggesting that loss of PagL function can occur during long-term adaptation to the CF airway.

* Corresponding author. Mailing address: Department of Medicine, University of Washington, HSB T-293, Box 357710, 1959 Pacific St. N.E., Seattle, WA 98195. Phone: (206) 616-4028. Fax: (206) 543-5383. E-mail: rkernst{at}u.washington.edu.

Journal of Bacteriology, January 2006, p. 191-201, Vol. 188, No. 1
0021-9193/06/$08.00+0     doi:10.1128/JB.188.1.191-201.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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