ENDOTOXIN BIOSYNTHESIS IN PSEUDOMONAS-AERUGINOSA - ENZYMATIC INCORPORATION OF LAURATE BEFORE 3-DEOXY-D-MANNO-OCTULOSONATE

被引:28
作者
MOHAN, S [1 ]
RAETZ, CRH [1 ]
机构
[1] MERCK & CO INC,RES LABS,DEPT BIOCHEM,RAHWAY,NJ 07065
关键词
D O I
10.1128/JB.176.22.6944-6951.1994
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Unlike Escherichia coli, living cells of Pseudomonas aeruginosa can complete the fatty acylation of lipid A when the biosynthesis of 3-deoxy-D-manno-octulosonate (Kdo) is inhibited (R. C. Goldman, C. C. Doran, S. K. Kadam, and J. O. Capobianco, J. Biol. Chem. 263:5217-5233, 1988). In this study, we demonstrate the presence of a novel enzyme in extracts of P. aeruginosa that can transfer laurate from lauroyl-acyl carrier protein (ACP) to a tetraacyl disaccharide-1,4'-bis-phosphate precursor of lipid A (termed lipid IVA) that accumulates in Kdo-deficient mutants off. coli. Comparable E. coli extracts cannot transfer laurate from lauroyl-ACP to lipid IVA, only to (Kdo)(2)-lipid IVA (K. A. Brozek, and C. R. H. Raetz, J. Biol. Chem. 265:15410-15517, 1990). P. aeruginosa extracts do not utilize myristoyl- or R-3-hydroxymyristoyl-ACP instead of lauroyl-ACP to acylate lipid IVA. Laurate incorporation in P. aeruginosa extracts is dependent upon time, protein concentration, and the presence of Triton X-100 but is inhibited by lauroyl-coenzyme A. P. aeruginosa extracts transfer only one laurate to lipid IVA, whereas E. coli extracts can transfer two laurates to (Kdo)(2)-lipid IVA. These results demonstrate that incorporation of laurate into lipid A does not require prior attachment of Kdo in all gram-negative bacteria.
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页码:6944 / 6951
页数:8
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