Uncommon membrane distribution of Shiga toxin glycosphingolipid receptors in toxin-sensitive human glomerular microvascular endothelial cells

Abstract Membrane microdomain association of the glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer), the highly and less effective receptors, respectively, for Shiga toxins (Stxs), is assumed as a functional requirement for Stx-mediated cytotoxicity. In a previous study, we demonstrated predominant localization of Stx receptors in cholesterol-enriched membrane microdomains of moderately Stx-sensitive human brain microvascular endothelial cells (HBMECs) by means of detergent-resistant membranes (DRMs). Here we report a different preferential distribution of Stx receptors in non-DRM fractions of human glomerular microvascular endothelial cells (GMVECs), the major targets of Stxs in the human kidney. Full structural characterization of Stx receptors using electrospray ionization (ESI) mass spectrometry revealed Gb3Cer and Gb4Cer lipoforms with ceramide moieties mainly composed of C24:0/C24:1 or C16:0 fatty acid and sphingosine (d18:1) in GMVECs comparable to those previously found in HBMECs. Thin-layer chromatography immunostaining demonstrated an approximately 2-fold higher content of Gb3Cer and a 1.4-fold higher content of Gb4Cer in GMVECs than in HBMECs. However, this does not explain the remarkable higher cytotoxic action of Stx1 and Stx2 toward GMVECs as compared with HBMECs. Our finding opens new questions on the microdomain association of Stx receptors and the functional role of GSLs in the membrane assembly of GMVECs.

[1]  K. Sandhoff,et al.  Mass spectrometric analysis of neutral sphingolipids: methods, applications, and limitations. , 2011, Biochimica et biophysica acta.

[2]  M. Mormann,et al.  Thin-layer chromatography, overlay technique and mass spectrometry: a versatile triad advancing glycosphingolipidomics. , 2011, Biochimica et biophysica acta.

[3]  A. Mellmann,et al.  Characterisation of the Escherichia coli strain associated with an outbreak of haemolytic uraemic syndrome in Germany, 2011: a microbiological study. , 2011, The Lancet. Infectious diseases.

[4]  S. Satchell,et al.  Acute laminar shear stress reversibly increases human glomerular endothelial cell permeability via activation of endothelial nitric oxide synthase , 2011, American journal of physiology. Renal physiology.

[5]  J. Rothberg,et al.  Prospective Genomic Characterization of the German Enterohemorrhagic Escherichia coli O104:H4 Outbreak by Rapid Next Generation Sequencing Technology , 2011, PloS one.

[6]  X. Wen,et al.  An In Vitro Model of the Glomerular Capillary Wall Using Electrospun Collagen Nanofibres in a Bioartificial Composite Basement Membrane , 2011, PloS one.

[7]  A. Edefonti,et al.  Clinical Relevance of Shiga Toxin Concentrations in the Blood of Patients With Hemolytic Uremic Syndrome , 2011, The Pediatric infectious disease journal.

[8]  A. Friedrich,et al.  Shiga toxin glycosphingolipid receptors in microvascular and macrovascular endothelial cells: differential association with membrane lipid raft microdomains[S] , 2011, Journal of Lipid Research.

[9]  N. Engedal,et al.  Toll-like receptor 4 facilitates binding of Shiga toxin to colon carcinoma and primary umbilical vein endothelial cells. , 2011, FEMS immunology and medical microbiology.

[10]  Patrik Langehanenberg,et al.  Differential cytotoxic actions of Shiga toxin 1 and Shiga toxin 2 on microvascular and macrovascular endothelial cells , 2010, Thrombosis and Haemostasis.

[11]  K. Sandvig,et al.  Endocytosis and retrograde transport of Shiga toxin. , 2010, Toxicon : official journal of the International Society on Toxinology.

[12]  P. Heeringa,et al.  Intrinsic renal cell and leukocyte-derived TLR4 aggravate experimental anti-MPO glomerulonephritis. , 2010, Kidney international.

[13]  T. Obrig.  Escherichia coli Shiga Toxin Mechanisms of Action in Renal Disease , 2010, Toxins.

[14]  P. Tazzari,et al.  Shiga toxin 1 and ricin A chain bind to human polymorphonuclear leucocytes through a common receptor. , 2010, The Biochemical journal.

[15]  A. Friedrich,et al.  On the structural diversity of Shiga toxin glycosphingolipid receptors in lymphoid and myeloid cells determined by nanoelectrospray ionization tandem mass spectrometry. , 2010, Rapid communications in mass spectrometry : RCM.

[16]  A. Friedrich,et al.  Neutral glycosphingolipids in human blood: a precise mass spectrometry analysis with special reference to lipoprotein-associated Shiga toxin receptors[S] , 2010, Journal of Lipid Research.

[17]  P. Boor,et al.  PDGF-C mediates glomerular capillary repair. , 2010, The American journal of pathology.

[18]  Tore-Geir Iversen,et al.  Protein toxins from plants and bacteria: Probes for intracellular transport and tools in medicine , 2010, FEBS letters.

[19]  B. Binnington,et al.  Globotriaosyl ceramide receptor function – Where membrane structure and pathology intersect , 2010, FEBS letters.

[20]  A. Surolia,et al.  Glycosphingolipids in microdomain formation and their spatial organization , 2010, FEBS letters.

[21]  J. Müthing,et al.  Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry. , 2010, Mass spectrometry reviews.

[22]  M. Morigi,et al.  Shiga toxin-associated hemolytic uremic syndrome: pathophysiology of endothelial dysfunction , 2010, Pediatric Nephrology.

[23]  J. Hunt.  Shiga Toxin–Producing Escherichia coli (STEC) , 2010, Clinics in Laboratory Medicine.

[24]  K. Sandvig,et al.  Interplay between Toxin Transport and Flotillin Localization , 2010, PloS one.

[25]  Kai Simons,et al.  Lipid Rafts As a Membrane-Organizing Principle , 2010, Science.

[26]  T. Okuda,et al.  Structural characterization and dynamics of globotetraosylceramide in vascular endothelial cells under TNF-α stimulation , 2010, Glycoconjugate Journal.

[27]  C. Sautès-Fridman,et al.  Hyperfunctional C3 convertase leads to complement deposition on endothelial cells and contributes to atypical hemolytic uremic syndrome. , 2009, Blood.

[28]  C. Lingwood,et al.  Fatty acid-dependent globotriaosyl ceramide receptor function in detergent resistant model membranes , 2009, Journal of Lipid Research.

[29]  A. Friedrich,et al.  Shiga Toxin Receptor Gb3Cer/CD77: Tumor-Association and Promising Therapeutic Target in Pancreas and Colon Cancer , 2009, PloS one.

[30]  Jeremy C. Allegood,et al.  Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers[S] , 2009, Journal of Lipid Research.

[31]  A. Shevchenko,et al.  Glycosphingolipid Requirements for Endosome‐to‐Golgi Transport of Shiga Toxin , 2009, Traffic.

[32]  C. Lingwood,et al.  Detergent-resistant globotriaosyl ceramide may define verotoxin/glomeruli-restricted hemolytic uremic syndrome pathology. , 2009, Kidney international.

[33]  Kazuro Furukawa,et al.  Immunologic glycosphingolipidomics and NKT cell development in mouse thymus. , 2009, Journal of proteome research.

[34]  S. Hell,et al.  Direct observation of the nanoscale dynamics of membrane lipids in a living cell , 2009, Nature.

[35]  S. Sonnino,et al.  Sphingolipids and membrane environments for caveolin , 2009, FEBS letters.

[36]  A. Friedrich,et al.  Shiga toxins, glycosphingolipid diversity, and endothelial cell injury , 2009, Thrombosis and Haemostasis.

[37]  S. Satchell,et al.  Shiga Toxin 2 Targets the Murine Renal Collecting Duct Epithelium , 2009, Infection and Immunity.

[38]  N. Loberto,et al.  Glycosphingolipid behaviour in complex membranes. , 2009, Biochimica et biophysica acta.

[39]  C. Sautès-Fridman,et al.  Hyperfunctional C 3 convertase leads to complement deposition on endothelial cells and contributes to atypical hemolytic uremic syndrome , 2009 .

[40]  K. Iwabuchi,et al.  Role of very long fatty acid-containing glycosphingolipids in membrane organization and cell signaling: the model of lactosylceramide in neutrophils , 2009, Glycoconjugate Journal.

[41]  A. Caprioli,et al.  Interactions between Shiga toxins and human polymorphonuclear leukocytes , 2008, Journal of leukocyte biology.

[42]  B. Binnington,et al.  Differential intracellular transport and binding of verotoxin 1 and verotoxin 2 to globotriaosylceramide‐containing lipid assemblies , 2008, Journal of cellular physiology.

[43]  A. Friedrich,et al.  Matching IR-MALDI-o-TOF mass spectrometry with the TLC overlay binding assay and its clinical application for tracing tumor-associated glycosphingolipids in hepatocellular and pancreatic cancer. , 2008, Analytical chemistry.

[44]  P. Wang,et al.  Sensitive quantitation of isoglobotriaosylceramide in the presence of isobaric components using electrospray ionization-ion trap mass spectrometry. , 2007, Glycobiology.

[45]  K. Murayama,et al.  Involvement of very long fatty acid-containing lactosylceramide in lactosylceramide-mediated superoxide generation and migration in neutrophils , 2008, Glycoconjugate Journal.

[46]  A. Friedrich,et al.  Glycosphingolipids in vascular endothelial cells: relationship of heterogeneity in Gb3Cer/CD77 receptor expression with differential Shiga toxin 1 cytotoxicity , 2008, Glycoconjugate Journal.

[47]  Katharina Gaus,et al.  Shiga toxin induces tubular membrane invaginations for its uptake into cells , 2007, Nature.

[48]  S. Satchell,et al.  Glomerular endothelial glycocalyx constitutes a barrier to protein permeability. , 2007, Journal of the American Society of Nephrology : JASN.

[49]  D. Lingwood,et al.  Detergent resistance as a tool in membrane research , 2007, Nature Protocols.

[50]  L. P. Van den Heuvel,et al.  Interactions of Shiga-like toxin with human peripheral blood monocytes , 2007, Pediatric Nephrology.

[51]  L. Johannes,et al.  Trafficking of Shiga toxin/Shiga-like toxin-1 in human glomerular microvascular endothelial cells and human mesangial cells. , 2006, Kidney international.

[52]  L. V. D. Heuvel,et al.  Conditionally immortalized human glomerular endothelial cells expressing fenestrations in response to VEGF. , 2006, Kidney international.

[53]  A. Friedrich,et al.  New aspects in the pathogenesis of enteropathic hemolytic uremic syndrome. , 2006, Seminars in thrombosis and hemostasis.

[54]  L. Johannes,et al.  Article Title: the Association of Shiga-like Toxin with Detergent-resistant Membranes Is Modulated by Glucosylceramide and Is an Essential Requirement in the Endoplasmic Reticulum for a Cytotoxic Effect the Association of Shiga-like Toxin with Detergent- Resistant Membranes Is Modulated by Glucosylc , 2022 .

[55]  W. Lencer,et al.  Raft trafficking of AB5 subunit bacterial toxins. , 2005, Biochimica et biophysica acta.

[56]  A. Friedrich,et al.  Application of combined high-performance thin-layer chromatography immunostaining and nanoelectrospray ionization quadrupole time-of-flight tandem mass spectrometry to the structural characterization of high- and low-affinity binding ligands of Shiga toxin 1. , 2005, Rapid communications in mass spectrometry : RCM.

[57]  H. Karch,et al.  Enterohaemorrhagic Escherichia coli in human medicine. , 2005, International journal of medical microbiology : IJMM.

[58]  H. Karch,et al.  Consequences of enterohaemorrhagic Escherichia coli infection for the vascular endothelium , 2005, Thrombosis and Haemostasis.

[59]  P. Tarr,et al.  Shiga-toxin-producing and haemolytic uraemic syndrome , 2005 .

[60]  B. Domon,et al.  A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates , 1988, Glycoconjugate Journal.

[61]  S. Levery.  Glycosphingolipid structural analysis and glycosphingolipidomics. , 2005, Methods in enzymology.

[62]  H. Dixon,et al.  IUPAC-IUB Joint Commission on Biochemical Nomenclature (JCBN) Nomenclature of unsaturated monosaccharides , 2005, Molecular and Cellular Biochemistry.

[63]  J. Peter-Katalinic,et al.  Direct analysis of silica gel extracts from immunostained glycosphingolipids by nanoelectrospray ionization quadrupole time-of-flight mass spectrometry. , 2004, Analytical chemistry.

[64]  J. Peter-Katalinic,et al.  Discrimination of neolacto-series gangliosides with alpha2-3- and alpha2-6-linked N-acetylneuraminic acid by nanoelectrospray ionization low-energy collision-induced dissociation tandem quadrupole TOF MS. , 2003, Analytical chemistry.

[65]  Deborah A. Brown,et al.  Use of Detergents to Study Membrane Rafts: The Good, the Bad, and the Ugly , 2003, Biological chemistry.

[66]  K. Sandhoff,et al.  Biosynthesis and degradation of mammalian glycosphingolipids. , 2003, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[67]  H. Karch,et al.  Cluster of hemolytic-uremic syndrome caused by Shiga toxin-producing Escherichia coli O26:H11 , 2003, The Pediatric infectious disease journal.

[68]  David G. E. Smith,et al.  Verotoxin 1 binding to intestinal crypt epithelial cells results in localization to lysosomes and abrogation of toxicity , 2003, Cellular microbiology.

[69]  A. Friedrich,et al.  Escherichia coli harboring Shiga toxin 2 gene variants: frequency and association with clinical symptoms. , 2002, The Journal of infectious diseases.

[70]  M. Donowitz,et al.  Role of lipid rafts in Shiga toxin 1 interaction with the apical surface of Caco-2 cells. , 2001, Journal of cell science.

[71]  D. Acheson,et al.  Shiga Toxin Translocation across Intestinal Epithelial Cells Is Enhanced by Neutrophil Transmigration , 2001, Infection and Immunity.

[72]  J. Salamero,et al.  Targeting of Shiga toxin B-subunit to retrograde transport route in association with detergent-resistant membranes. , 2001, Molecular biology of the cell.

[73]  R. Schnaar,et al.  Segregation of Gangliosides GM1 and GD3 on Cell Membranes, Isolated Membrane Rafts, and Defined Supported Lipid Monolayers , 2001, Biological chemistry.

[74]  K. Sandvig.  Shiga toxins. , 2001, Toxicon : official journal of the International Society on Toxinology.

[75]  D. Smith,et al.  Toxin entry: how bacterial proteins get into mammalian cells , 1999, Cellular microbiology.

[76]  J. Peter-Katalinic,et al.  Isolation and structural characterization of glycosphingolipids of in vitro propagated human umbilical vein endothelial cells. , 1999, Glycobiology.

[77]  M.Alan Chester,et al.  IUPAC-IUB Joint Commission on Biochemical Nomenclature (JCBN). Nomenclature of glycolipids--recommendations 1997. , 1998, European journal of biochemistry.

[78]  D. Welch.  Cat-scratch disease; Negative urine dipstick, positive culture; Oryx initiative , 1999 .

[79]  J. Müthing.  TLC in structure and recognition studies of glycosphingolipids. , 1998, Methods in molecular biology.

[80]  F. Gilles,et al.  Selective expression of adhesion molecules on human brain microvascular endothelial cells , 1997, Journal of Neuroimmunology.

[81]  C. Lingwood.  Role of verotoxin receptors in pathogenesis. , 1996, Trends in microbiology.

[82]  Jeanette Adams,et al.  Structure determination of sphingolipids by mass spectrometry , 1993 .

[83]  Deborah A. Brown,et al.  Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface , 1992, Cell.

[84]  R. Cotter,et al.  Mass Spectrometry , 1992, Bio/Technology.

[85]  B. Domon,et al.  Structure elucidation of glycosphingolipids and gangliosides using high-performance tandem mass spectrometry. , 1988, Biochemistry.

[86]  T. Yamakawa,et al.  Structural study on tetraglycosyl ceremide and gangliosides isolated from human red blood cells. , 1973, Journal of biochemistry.

[87]  C. Frey,et al.  Manganous chloride spray reagent for cholesterol and bile acids on thin-layer chromatograms. , 1970, Journal of chromatography.

[88]  S. Siegel,et al.  Nonparametric Statistics for the Behavioral Sciences , 2022, The SAGE Encyclopedia of Research Design.

[89]  E. G. Olds.  Distributions of Sums of Squares of Rank Differences for Small Numbers of Individuals , 1938 .