Identification of Staphylococcal Enterotoxin B Domains Involved in Binding to Cultured Human Kidney Proximal Tubular Cells: Imparting Proliferation and Death

Abstract

Studies suggest that staphylococcal enterotoxin B (SEB) is initially harbored in the kidney by binding to digalactosylceramide molecules in the proximal tubular cells. However, little is known in regard to the peptide motif within SEB that binds to these cells and imparts toxic effects. Herein, using human kidney proximal tubular cells (PTs) we have performed a systematic study on the binding of various peptides and peptide analogs of SEB and demonstrate a structure-functional relationship. Using [¹²⁵I]labeled SEB peptides, we show a high affinity and displaceable binding of SEB 191–220 to human PT cells. Binding was mitigated by the use of antibody against SEB, by digalactosylceramide (the putative receptor), and by the use of endoglycoceramidase, which selectively removes the oligosaccharide backbones from glycosphingolipids. Our structure/ functional studies revealed that peptide 130–160 induces a concentration-dependent increase in programmed cell death/ apoptosis in human proximal tubular cells. Mechanistic studies further suggest that SEB/SEB peptide (130–160) impart apoptosis via the activation of neutral sphingomyelinase, which hydrolizes sphingomyelin to ceramide and phosphocholine. SEB 130–160 mediated apoptosis was mitigated by preincubation of cells with antibody against SEB and an SEB 130–160 antibody.

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References

Jett M, Ionin B, Das R, Neill R. The staphylococcal enterotoxins. In: Sussman M, Ed. Molecular Medical Microbiology. San Diego: Academic Press, 2001.

Jardetzky TS, Brown JH, Gorga JC, Stern LJ, Urban RG, Chi YI, Stauffacher C, Strominger JL, Wiley DC. Three-dimensional structure of a human class II histocompatibility molecule complexed with superantigen. Nature 368:711–718, 1994.

Kappler JW, Herman A, Clements J, Marrack P. Mutations defining functional regions of the superantigen staphylococcal enterotoxin B. J Exp Med 175:387–396, 1992.

Hoffman M, Tremaine M, Mansfield J, Betley M. Biochemical and mutational analysis of the histidine residues of staphylococcal enterotoxin A. Infect Immun 64:885–890, 1996.

Spero L, Morlock BA. Biological activities of the peptides of staphylococcal enterotoxin C formed by limited tryptic hydrolysis. J Biol Chem 253:8787–8791, 1978.

Campbell WN, Fitzpatrick M, Ding X, Jett M, Gemski P, Goldblum SE. SEB is cytotoxic and alters EC barrier function through protein tyrosine phosphorylation in vitro. Am J Physiol 273:L31–L39, 1997.

Shupp JW, Jett M, Pontzer CH. Identification of a transcytosis epitope on staphylococcal enterotoxins. Infect Immun 70:2178–2186, 2002.

van Gessel YA, Mani S, Bi S, Hammamieh R, Shupp JW, Das R, Coleman GD, Jett M. Functional piglet model for the clinical syndrome and postmortem findings induced by staphylococcal enterotoxin B. Exp Biol Med (Maywood) 229:1061–1071, 2004.

Canonico PG, Henriksen EL, Ayala E, Bowman DG. Subcellular localization of staphylococcal enterotoxin B in rabbit kidney. Am J Physiol 226:1333–1337, 1974.

10.

Morris EL, Hodoval LF, Beisel WR. The unusual role of the kidney during intoxication of monkeys by intravenous staphylococcal enterotoxin B. J Infect Dis 117:273–284, 1967.

11.

Normann SJ, Jaeger RF, Johnsey RT. Pathology of experimental enterotoxemia. The in vivo localization of staphylococcal enterotoxin B. Lab Invest 20:17–25, 1969.

12.

Normann SJ, Stone CM. Renal lysosomal catabolism of staphylococcal enterotoxin B. Lab Invest 27:236–241, 1972.

13.

Papageorgiou AC, Tranter HS, Acharya KR. Crystal structure of microbial superantigen staphylococcal enterotoxin B at 1.5 A resolution: implications for superantigen recognition by MHC class II molecules and T-cell receptors. J Mol Biol 277:61–79, 1998.

14.

Rapoport MI, Hodoval LF, Beisel WR. Influence of thorotrast blockade and acute renal artery ligation on disappearance of staphylococcal enterotoxin B from blood. J Bacteriol 93:779–783, 1967.

15.

Chatterjee S, Jett M. Glycosphingolipids: the putative receptor for Staphylococcus aureus enterotoxin-B in human kidney proximal tubular cells. Mol Cell Biochem 113:25–31, 1992.

16.

Chatterjee S, Khullar M, Shi WY. Digalactosylceramide is the receptor for staphylococcal enterotoxin-B in human kidney proximal tubular cells. Glycobiology 5:327–333, 1995.

17.

Chatterjee S, Sekerke CS, Kwiterovich PO Jr. Increased urinary excretion of glycosphingolipids in familial hypercholesterolemia. J Lipid Res 23:513–522, 1982.

18.

Chatterjee S, Kwiterovich PO. Glycosphingolipids of human plasma lipoproteins. Lipids. 11:462–466, 1976.

19.

Markwell MA, Fox CF. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry 17:4807–4817, 1978.

20.

Chatterjee S, Kwiterovich PO Jr, Gupta P, Erozan YS, Alving CR, Richards RL. Localization of urinary lactosylceramide in cytoplasmic vesicles of renaltubular cells in homozygous familial hypercholesterolemia. Proc Natl Acad Sci U S A 80:1313–1317, 1983.

21.

Jett M, Neill R, Welch C, Boyle T, Bernton E, Hoover D, Lowell G, Hunt RE, Chatterjee S, Gemski P. Identification of staphylococcal enterotoxin B sequences important for induction of lymphocyte proliferation by using synthetic peptide fragments of the toxin. Infect Immun 62:3408–3415, 1994.

22.

Chatterjee S. Neutral sphingomyelinase: past, present and future. Chem Phys Lipids 102:79–96, 1999.

23.

Obeid LM, Linardic CM, Karolak LA, Hannun YA. Programmed cell death induced by ceramide. Science 259:1769–1771, 1993.

24.

Luberto C, Yoo DS, Suidan HS, Bartoli GM, Hannun YA. Differential effects of sphingomyelin hydrolysis and resynthesis on the activation of NF-kappa B in normal and SV40-transformed human fibroblasts. J Biol Chem 275:14760–14766, 2000.

25.

Lycke N, Bromander AK, Ekman L, Karlsson U, Holmgren J. Cellular basis of immunomodulation by cholera toxin in vitro with possible association to the adjuvant function in vivo. J Immunol 142:20–27, 1989.

26.

Chatterjee S, Martin S. Sphingolipid metabolism and signaling in atherosclerosis. Adv Cell Aging Ger 12:71–96, 2003.

27.

Spiegel S, Matyas GR, Cheng L, Sacktor B. Asymmetric distribution of gangliosides in rat renal brush-border and basolateral membranes. Biochim Biophys Acta 938:270–278, 1988.