Abstract
Lipopolysaccharide (LPS)-binding protein (LBP) is a critical component of innate immunity, implicated in the initiation of host defences against Gram-negative bacteria. LBP alerts the host to the presence of minute amounts of LPS (1). LPS released from Gram-negative bacteria is present as aggregates, because of the amphiphilic structure of the molecule. LPS aggregates are transformed to monomers by the action of LBP, which has been described as a lipid-transfer molecule catalyzing movement of phospholipids including LPS (2–6). When LPS/LBP monomers are transferred to lipoproteins, LPS is inactivated; when LPS/LBP complexes are transferred to cells harboring CD14 at their surface, cells are activated. Thus, the relative contribution of these two pathways will determine the response of the host to LPS.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Schumann, R. R., Leong, S. R., Flaggs, G. W., Gray, P. W., Wright, S. D., Mathison, J. C., Tobias, P. S., and Ulevitch, R. J. (1990) Structure and function of lipopolysaccharide binding protein. Science 249, 1429–1431.
Wurfel, M. M., Hailman, E., and Wright, S. D. (1995) Soluble CD14 acts as a shuttle in the neutralization of lipopolysaccharide (LPS) by LPS-binding protein and reconstituted high density lipoprotein. J. Exp. Med. 181, 1743–1754.
Yu, B., Hailman, E., and Wright, S. D. (1997) Lipopolysaccharide binding protein and soluble CD14 catalyze exchange of phospholipids. J. Clin. Invest. 99, 315–324.
Park, C. T. and Wright, S. D. (1996) Plasma lipopolysaccharide-binding protein is found associated with a particle containing apolipoprotein A-I, phospholipid, and factor H-related proteins. J. Biol. Chem. 271, 18,054–18,060.
Wurfel, M. M. and Wright, S. D. (1997) Lipopolysaccharide-binding protein and soluble CD14 transfer lipopolysaccharide to phospholipid bilayers. Preferential interaction with particular classes of lipid. J. Immunol. 158, 3925–3934.
Yu, B. and Wright, S. D. (1996) Catalytic properties of lipopolysaccharide (LPS) binding protein. J. Biol. Chem. 271, 4100–4105.
Gallay, P., Heumann, D., Le Roy, D., Barras, C., and Glauser, M. P. (1993) Lipopolysaccharide-binding protein as a major plasma protein responsible for endotoxemic shock. Proc. Natl. Acad. Sci. USA 90, 9935–9938.
Gallay, P., Heumann, D., Le Roy, D., Barras, C., and Glauser, M. P. (1994) Mode of action of anti-lipopolysaccharide binding protein (LBP) antibodies for prevention of endotoxemic shock in mice. Proc. Natl. Acad. Sci. USA 91, 7922–7926.
Jack, R. S., Fan, X. L., Bernheiden, M., Rune, G., Ehlers, M., Weber, A., Kirsch, G., Mentel, R., Furll, B., Freudenberg, M., Schmitz, G., Stelter, F., and Schütt, C. (1997) Lipopolysaccharide-binding protein is required to combat a Gram-negative bacterial infection. Nature 389, 742–745.
Lengacher, S., Jongeneel, C. V., Le Roy, D., Lee, J. D., Kravtchenko, V., Ulevitch, R. J., Glauser, M. P., and Heumann, D. (1996) Reactivity of murine and human recombinant LPS-binding protein (LBP) with LPS and Gram-negative bacteria. J. Inflamm. 47, 165–172.
Goldblum, S. E., Brann, T. W., Ding, X., Pugin, J., and Tobias, P. S. (1994) Lipopolysaccharide (LPS)-binding protein and soluble CD14 function as accesory molecules for LPS-induced changes in endothelial barrier function, in vitro. J. Clin. Invest. 93, 692–702.
Lee, J. D., Kato, K., Tobias, P. S., Kirkland, T. N., and Ulevitch, R. J. (1992) Transfection of CD14 into 70Z/3 cells dramatically enhances the sensitivity to complexes of lipopolysaccharide (LPS) and LPS binding protein. J. Exp. Med. 175, 1697–1705.
Lee, J. D., Kravchenko, V., Kirkland, T. N., Han, J., Mackman, N., Moriarty, A., Leturcq, D., Tobias, P. S., and Ulevitch, R. J. (1993) Glycosyl-phosphatidyl-anchored or integral membrane forms of CD14 mediate identical cellular responses to endotoxin. Proc. Natl. Acad. Sci. USA 90, 9930–9934.
Tobias, P. S., Soldau, K., Gegner, J. A., Mintz, D., and Ulevitch, R. J. (1995) Lipopolysaccharide binding protein-mediated complexation of lipopolysaccharide with soluble CD14. J. Biol. Chem. 270, 10,482–10,488.
Delude, R. L., Fenton, M. J., Jr., Savedra, R., Perera, P. Y., Vogel, S. N., Thieringer, R., and Golenblock, D. T. (1994) CD14-mediated translocation of nuclear factor-κB induced by lipopolysaccharide does not require tyrosine kinase activity. J. Biol. Chem. 269, 22,253–22,260.
Le Roy, D., Di Padova, F., Tees, R., Lengacher, S., Landmann, R., Glauser, M. P., Calandra, T., and Heumann, D. (1999) Monoclonal antibodies to murine lipopolysaccharide (LPS)-binding protein (LBP) protect mice from lethal endotoxemia by blocking either the binding of LPS to LPB or the presentation of LPS/LBP complexes to CD14. Journal of Immunology 162, 7454–7460.
Skelly, R. R., Munkenbeck, P., and Morrison, D. C. (1979) Stimulation of T-independent antibody responses by hapten-lipopolysaccharides without repeating polymeric structure. Infect.Immun. 23, 287–293.
Heumann, D., Gallay, P., Barras, C., Zaech, P., Ulevitch, R. J., Tobias, P. S., Glauser, M. P., and Baumgartner, J. D. (1992) Control of LPS binding and LPS-induced TNF secretion in human peripheral blood monocytes. J. Immunol. 148, 3505–3512.
Tobias, P. S., Soldau, K., and Ulevitch, R. J. (1986) Isolation of a lipopolysaccharide-binding acute phase reactant from rabbit serum. J. Exp. Med. 164, 777–793.
Gallay, P., Carrel, S., Glauser, M. P., Barras, C., Ulevitch, R. J., Tobias, P. S., Baumgartner, J. D., and Heumann, D. (1993) Purification and characterization of murine LPS binding protein. Infect. Immun. 61, 378–383.
Khemlani, L. S., Yang, Z., and Bochsler, P. N. (1994) Identification and characterization of a bovine lipopolysaccharide-binding protein. J. Leuk. Biol. 56, 784–791.
Theofan, G., Horwitz, A. H., Williams, R. E., Liu, P. S., Chan, I., Birr, C., Carroll, S. F., Meszaros, K., Parent, J. B., Kasler, H., Aberle, S., Trown, P. W., and Gazzano-Santoro, H. (1994) An amino-terminal fragment of human lipopolysaccharide-binding protein retains lipid A binding but not CD14-stimulatory activity. J. Immunol. 152, 3623–3629.
Heumann, D., Gallay, P., Le Roy, D., and Glauser, M. P. (1994) Radioimmunoassay versus flow cytometric assay to quantify LPS-binding protein (LBP) concentrations in human plasma. J. Immunol. Methods 171, 169–176.
Dubin, W., Martin, T. R., Swoveland, P., Leturcq, D. J., Moriarty, A. M., Tobias, P. S., Bleicker, C. A., Goldblum, S. E., and Hasday, J. D. (1996) Asthma and endotoxin: lipopolysaccharide-binding protein and soluble CD14 in bronchoalveolar compartment. Am. J. Physiol. 270(5 Pt1), L736–L744.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Heumann, D. (2000). Purification of Lipopolysaccharide Binding Protein. In: Evans, T.J. (eds) Septic Shock Methods and Protocols. Methods in Molecular Medicine™, vol 36. Humana Press. https://doi.org/10.1385/1-59259-216-3:45
Download citation
DOI: https://doi.org/10.1385/1-59259-216-3:45
Publisher Name: Humana Press
Print ISBN: 978-0-89603-730-4
Online ISBN: 978-1-59259-216-6
eBook Packages: Springer Protocols