Complement attack of altered outer membrane areas synthesized after inhibition of the 3-deoxy-D-manno-octulosonate pathway leads to cell death.
Salmonella typhimurium containing specific genes coding for either temperature-sensitive (TS) 3-deoxy-D-manno-octulosonate (KDO) 8-phosphate synthetase or TS cytidine monophosphate-KDO synthetase grow normally when incubated at 30 degrees C and are resistant to C-mediated killing. However, bacteria become avirulent and sensitive to C-mediated killing upon thermal inhibition of TS KDO-8-phosphate synthetase (incubation at 38 degrees C) or TS cytidine monophosphate-KDO synthetase (incubation at 42 degrees C). Such thermal inhibition concurrently causes synthesis of an altered outer membrane which we now show is the site that renders cells susceptible to C-mediated killing. After incubation of cells in serum, the altered outer membrane area contains C9 in a trypsin-resistant state and membrane attack complex (MAC) lesions observable by electron microscopy. Trypsin-resistant C9 and MAC lesions were also observed in the inner membrane fraction from such serum-treated cells. In contrast, little C9 and few MAC lesions were associated with unaltered outer membrane areas present on these same serum treated cells. Control cells, grown at 30 degrees C and treated with serum (1) bound one-fifth as much C9 as was bound to cells incubated at 42 degrees C, (2) contained only a rare MAC lesion in the outer membrane, and (3) no observable MAC lesions in the inner membrane. We conclude that the altered outer membrane area is the site that renders cells susceptible to insertion of the MAC into both the outer and inner membrane resulting in cell death.
Goldman
R C
RC
Anti-Infective Research Division, Abbott Laboratories, Abbott Park, IL 60064.
Miller
M F
MF
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000648:ultrastructure; D001770:Blood Bactericidal Activity; D015153:Blotting, Western; D003186:Complement C9 / Q000378:metabolism; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D005260:Female; D006017:Glycolipids / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D008050:Lipid A / Q000031:analogs & derivatives / Q000378:metabolism; D051379:Mice; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors*; D011951:Receptors, Complement / Q000032:analysis; D012486:Salmonella typhimurium / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure
D001425:Bacterial Outer Membrane Proteins; D003186:Complement C9; D015938:Complement Membrane Attack Complex; D006017:Glycolipids; D008050:Lipid A; D011951:Receptors, Complement; C040142:lipid A precursors, bacterial; D003165:Complement System Proteins; D009713:Nucleotidyltransferases; C028984:3-deoxy-manno-octulosonate cytidyltransferase