PMID: 9428693Jan 15, 1998Paper

Detection of lipopolysaccharide (LPS)-binding membrane proteins by immuno-coprecipitation with LPS and anti-LPS antibodies

European Journal of Biochemistry
V T El-SamaloutiA J Ulmer

Abstract

In this study we describe a general method for the detection and characterization of endotoxin-(lipopolysaccharide, LPS)-binding membrane proteins. In the past, experimental procedures to detect LPS-binding sites on cells were generally performed with chemically modified LPS derivates. Since any modification of a ligand may lead to a modification of its binding characteristics, the results of those studies are controversial. In our assay, cell membrane preparations are treated with free lipid A, the endotoxic center of LPS, in the presence of normal human serum. After binding of lipid A, membrane proteins are solubilized by mild detergent treatment without disruption of the lipid A-protein complexes. Addition of anti-(lipid A) mAbs and subsequent adding of protein A agarose lead to the precipitation of complexes of lipid A and its binding proteins. By SDS/PAGE and western blot, these precipitates can be screened for the presence of LPS/lipid A-binding proteins. We describe the use of this method for the immuno-coprecipitation of lipid A (or LPS) with an 80-kDa LPS-binding membrane protein (LMP80), which we have previously identified on several human cells. In addition, CD14, the well-known functional LPS receptor on monocytes a...Continue Reading

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Citations

Jun 20, 2002·Trends in Immunology·Martha Triantafilou, Kathy Triantafilou
Sep 24, 1999·Biological Chemistry·A WieseS Müller-Loennies
Jul 27, 2010·Analytical and Bioanalytical Chemistry·Carlo BertucciAngela De Simone
Aug 23, 2002·The Journal of Investigative Dermatology·Peter I SongJohn C Ansel
Mar 22, 2015·Seminars in Nephrology·Sudhanshu Kumar Verma, Bruce A Molitoris
Nov 24, 2011·Toxicologic Pathology·Wen-Chao Song
Jun 19, 2015·The Journal of Biological Chemistry·Omid Haji-GhassemiStephen V Evans
Mar 5, 2016·The Journal of Biological Chemistry·Omid Haji-GhassemiStephen V Evans
Jan 29, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yasuhiko YamamotoHiroshi Yamamoto
Apr 29, 1999·FEMS Immunology and Medical Microbiology·V T el-SamaloutiL Hamann

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