Proteins of 30 and 36 kilodaltons, membrane constituents of the Staphylococcus aureus L form, induce production of tumor necrosis factor alpha and activate the human immunodeficiency virus type 1 long terminal repeat.
We have previously demonstrated that the membrane of the Staphylococcus aureus L form induced tumor necrosis factor alpha (TNF-alpha) from murine macrophages. In this study, we purified two proteins which induce TNF-alpha production from a human monocytic cell line, THP-1, and murine macrophages. These molecules were purified from delipidated membranes by deoxycholic acid extraction, two-step anion-exchange chromatography, and preparative electrophoresis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified proteins showed for each a single band with a molecular mass of 30, and 36 kDa. These proteins were heat stable. Polymyxin B did not affect the production of TNF-alpha induced by these proteins. Furthermore, these proteins induced comparable levels of TNF-alpha in both lipopolysaccharide-responsive and -nonresponsive mouse macrophages. Pretreatment of murine macrophages with gamma interferon enhanced 30- and 36-kDa protein-mediated TNF-alpha production. The 30-kDa protein showed lethal toxicity to D-galactosamine-treated mice. The 30- and 36-kDa proteins stimulated the human immunodeficiency virus type 1 long terminal repeat in a monocytic cell line but not a T-cell line. This effect appeared to be mediated through the induction of nuclear factor kappaB. These results indicate that the 30- and 36-kDa proteins, membrane constituents of the S. aureus L form, may play a role in S. aureus infection and/or in human immunodeficiency virus type 1-infected individuals.
Akashi
A
A
Department of Microbiology, Kurume University School of Medicine, Japan.
Ono
S
S
Kuwano
K
K
Arai
S
S
D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D015967:Gene Expression Regulation, Viral; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D007740:L Forms / Q000737:chemistry*; D008262:Macrophage Activation; D017737:Macrophages, Peritoneal / Q000187:drug effects; D008565:Membrane Proteins / Q000302:isolation & purification / Q000494:pharmacology* / Q000633:toxicity; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D009000:Monocytes / Q000821:virology; D011112:Polymyxin B / Q000494:pharmacology; D011489:Protein Denaturation; D013211:Staphylococcus aureus / Q000737:chemistry*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D001426:Bacterial Proteins; D008565:Membrane Proteins; D014409:Tumor Necrosis Factor-alpha; D011112:Polymyxin B