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© The Rockefeller University Press, 0022-1007/1999/3/907/ $5.00
The Journal of Experimental Medicine, Volume 189, Number 6, March 15, 1999 907-918

Articles

An Inflammatory Polypeptide Complex from Staphylococcus epidermidis: Isolation and Characterization

Christopher Mehlin{ddagger}, Catherine M. Headley*, and Seymour J. Klebanoff*,{ddagger}

From the * Department of Medicine and the {ddagger} Department of Pathobiology, University of Washington, Seattle, Washington 98195

Staphylococcus epidermidis releases factors that activate the HIV-1 long terminal repeat, induce cytokine release, and activate nuclear factor {kappa}B in cells of macrophage lineage. The active material had a mass of 34,500 daltons, was inactivated by proteases and partitioned into the phenol layer on hot aqueous phenol extraction, and thus was termed phenol-soluble modulin (PSM). High performance liquid chromatography (HPLC) of crude PSM yielded two peaks of activity designated PSM peak 1 and peak 2. MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) mass spectroscopy indicated the presence of two components in peak 1, which were designated PSM{alpha} and PSMβ. Peak 2 contained a single component, designated PSM{gamma}. Separation of PSM{alpha} and PSMβ in peak 1 could be achieved by a second HPLC procedure. The structure of each component was determined by amino acid sequence analysis and identification and sequencing of their genes. PSM{alpha}, PSMβ, and PSM{gamma} were 22-, 44-, and 25-amino acid, respectively, strongly hydrophobic polypeptides. PSM{gamma} was identified as Staphylococcus epidermidis delta toxin, whereas PSM{alpha} and PSMβ exhibited more distant homology to previously described staphylococcal toxins. They appeared to exist as a complex or aggregate with activity greater than the component parts. The properties of the S. epidermidis PSMs suggest that they may contribute to the systemic manifestations of Gram-positive sepsis.

Key Words: Staphylococcus epidermidis • HIV-1 long terminal repeat • phenol-soluble modulin • inflammatory polypeptide • nuclear factor {kappa}B

Address correspondence to Seymour J. Klebanoff, Department of Medicine, Box 357185, University of Washington, Seattle, WA 98195. Phone: 206-543-7902; Fax: 206-685-8681; E-mail: seym{at}u.washington.edu

Abbreviations used: LTA, lipoteichoic acid; MALDI-TOF, matrix-assisted laser desorption ionization-time of flight; MWCO, molecular weight cut off; NEC, neonatal necrotizing enterocolitis; PSM, phenol-soluble modulin; RLU, relative light units.


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