A novel Escherichia coli lipid A mutant that produces an antiinflammatory lipopolysaccharide.
A unique screen was used to identify mutations in Escherichia coli lipid A biosynthesis that result in a decreased ability to stimulate E-selectin expression by human endothelial cells. A mutation was identified in the msbB gene of E. coli that resulted in lipopolysaccharide (LPS) that lacks the myristoyl fatty acid moiety of the lipid A. Unlike all previously reported lipid A mutants, the msbB mutant was not conditionally lethal for growth. Viable cells or purified LPS from an msbB mutant had a 1000-10,000-fold reduction in the ability to stimulate E-selectin production by human endothelial cells and TNF alpha production by adherent monocytes. The cloned msbB gene was able to functionally complement the msbB mutant, restoring both the LPS to its native composition and the ability of the strain to stimulate immune cells. Nonmyristoylated LPS acted as an antagonist for E-selectin expression when mixed with LPS obtained from the parental strain. These studies demonstrate a significant role for the myristate component of LPS in immune cell activation and antagonism. In addition, the msbB mutant allowed us to directly examine the crucial role that the lipid A structure plays when viable bacteria are presented to host defense cells.
Somerville
J E
JE
Inflammation Department, Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121, USA.
Cassiano
L
L
Bainbridge
B
B
Cunningham
M D
MD
Darveau
R P
RP
D000217:Acyltransferases; D002478:Cells, Cultured; D019040:E-Selectin / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology*; D029968:Escherichia coli Proteins; D005227:Fatty Acids / Q000737:chemistry; D006801:Humans; D007535:Isomerases / Q000235:genetics; D008050:Lipid A / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000737:chemistry / Q000502:physiology; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000737:chemistry; D016296:Mutagenesis; D019814:Myristic Acid; D009227:Myristic Acids / Q000737:chemistry; D019704:Protein Disulfide-Isomerases; D013329:Structure-Activity Relationship; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D019040:E-Selectin; D029968:Escherichia coli Proteins; D005227:Fatty Acids; D008050:Lipid A; D008070:Lipopolysaccharides; D009227:Myristic Acids; D014409:Tumor Necrosis Factor-alpha; D019814:Myristic Acid; D000217:Acyltransferases; C097895:MsbB protein, E coli; D007535:Isomerases; D019704:Protein Disulfide-Isomerases