Mouse anti-ceramide antiserum: a specific tool for the detection of endogenous ceramide.

Abstract:

:Ceramide is a pivotal molecule in signal transduction and an essential structural component of the epidermal permeability barrier. The epidermis is marked by a high concentration of ceramide and by a unique spectrum of ceramide species: Besides the two ceramide structures commonly found in mammalian tissue, N-acylsphingosine and N-2-hydroxyacyl-sphingosine, six additional ceramides differing in the grade of hydroxylation of either the sphingosine base or the fatty acid have been identified in the epidermis. Here we report on the characterization of an IgM-enriched polyclonal mouse serum against ceramide. In dot blot assays with purified epidermal lipids the antiserum bound to a similar extent to N-acyl-sphingosine (ceramide 2), N-acyl-4-hydroxysphinganine (ceramide 3), and N-(2-hydroxyacyl)-sphingosine (ceramide 5), whereas no specific reaction was detected with glycosylceramides, sphingomyelin, free sphingosine, phospholipids, or cholesterol. In contrast, a monoclonal IgM antibody, also claimed to be specific for ceramide, was shown to bind specifically to sphingomyelin and therefore was not further investigated. In thin-layer chromatography immunostaining with purified lipids a strong and highly reproducible reaction of the antiserum with ceramide 2 and ceramide 5 was observed, whereas the reaction with ceramide 1 and ceramide 3 was weaker and more variable. Ceramide 2 and ceramide 5 were detected in the nanomolar range at serum dilutions of up to 1:100 by dot blot and thin-layer immunostaining. In thin-layer chromatography immunostaining of crude lipid extracts from human epidermis, the antiserum also reacted with N-(2-hydroxyacyl)-4-hydroxysphinganine (ceramide 6) and N-(2-hydroxyacyl)-6-hydroxysphingosine (ceramide 7). Furthermore, the suitability of the antiserum for the detection of endogenous ceramide by immunolight microscopy was demonstrated on cryoprocessed human skin tissue. Double immunofluorescence labeling experiments with the anti-ceramide antiserum and the recently described anti-glucosylceramide antiserum (Brade et al., 2000, Glycobiology 10, 629) showed that both lipids are concentrated in separate epidermal sites. Whereas anti-ceramide stained the dermal and basal epidermal cells as well as the corneocytes, anti-glucosylceramide staining was concentrated in the stratum granulosum. In conclusion, the specificity and sensitivity of the reagent will enable studies on the subcellular distribution and biological functions of endogenous ceramide.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Vielhaber G,Brade L,Lindner B,Pfeiffer S,Wepf R,Hintze U,Wittern KP,Brade H

doi

10.1093/glycob/11.6.451

subject

Has Abstract

pub_date

2001-06-01 00:00:00

pages

451-7

issue

6

eissn

0959-6658

issn

1460-2423

journal_volume

11

pub_type

杂志文章
  • Cryptococcus neoformans UGT1 encodes a UDP-Galactose/UDP-GalNAc transporter.

    abstract::Cryptococcus neoformans, an opportunistic fungal pathogen, produces a glycan capsule to evade the immune system during infection. This definitive virulence factor is composed mainly of complex polysaccharides, which are made in the secretory pathway by reactions that utilize activated nucleotide sugar precursors. Alth...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cww078

    authors: Li LX,Ashikov A,Liu H,Griffith CL,Bakker H,Doering TL

    更新日期:2017-01-01 00:00:00

  • Characterization of N-linked oligosaccharides assembled on secretory recombinant glucose oxidase and cell wall mannoproteins from the methylotrophic yeast Hansenula polymorpha.

    abstract::Presently almost no information is available on the oligosaccharide structure of the glycoproteins secreted from the methylotrophic yeast Hansenula polymorpha, a promising host for the production of recombinant proteins. In this study, we analyze the size distribution and structure of N-linked oligosaccharides attache...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh030

    authors: Kim MW,Rhee SK,Kim JY,Shimma Y,Chiba Y,Jigami Y,Kang HA

    更新日期:2004-03-01 00:00:00

  • Molecular characterization of the cis-prenyltransferase of Giardia lamblia.

    abstract::Giardia lamblia, the protist that causes diarrhea, makes an Asn-linked-glycan (N-glycan) precursor that contains just two sugars (GlcNAc(2)) attached by a pyrophosphate linkage to a polyprenol lipid. Because the candidate cis-prenyltransferase of Giardia appears to be more similar to bacterial enzymes than to those of...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwq036

    authors: Grabińska KA,Cui J,Chatterjee A,Guan Z,Raetz CR,Robbins PW,Samuelson J

    更新日期:2010-07-01 00:00:00

  • A novel sialylated N-acetylgalactosamine-containing oligosaccharide is the major complex-type structure present in Bowes melanoma tissue plasminogen activator.

    abstract::We have employed fast atom bombardment mass spectrometry (FAB-MS) to screen the N-linked oligosaccharides of Bowes melanoma tissue plasminogen activator (mt-PA), and recombinant t-PAs produced by Chinese hamster ovary cells (rt-PA) and by a gene-enriched melanoma cell line (rmt-PA). These studies have confirmed the pu...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/1.2.173

    authors: Chan AL,Morris HR,Panico M,Etienne AT,Rogers ME,Gaffney P,Creighton-Kempsford L,Dell A

    更新日期:1991-03-01 00:00:00

  • Isolation and characterization of a class II alpha-mannosidase cDNA from lepidopteran insect cells.

    abstract::Lepidopteran insect cells are used routinely as hosts for foreign glycoprotein expression by recombinant baculoviruses, but the precise nature of their N-glycosylation pathway remains poorly defined. These cells clearly have processing glucosidases and mannosidases that can convert precursors to Man3GlcNAc2 structures...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/7.1.113

    authors: Jarvis DL,Bohlmeyer DA,Liao YF,Lomax KK,Merkle RK,Weinkauf C,Moremen KW

    更新日期:1997-02-01 00:00:00

  • Genetic basis for expression of the major globotetraose-containing lipopolysaccharide from H. influenzae strain Rd (RM118).

    abstract::A genetic basis for the biosynthetic assembly of the globotetraose containing lipopolysaccharide (LPS) of Haemophilus influenzae strain RM118 (Rd) was determined by structural analysis of LPS derived from mutant strains. We have previously shown that the parent strain RM118 elaborates a population of LPS molecules mad...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/11.11.957

    authors: Hood DW,Cox AD,Wakarchuk WW,Schur M,Schweda EK,Walsh SL,Deadman ME,Martin A,Moxon ER,Richards JC

    更新日期:2001-11-01 00:00:00

  • Specificity of IgG and IgE antibodies against plant and insect glycoprotein glycans determined with artificial glycoforms of human transferrin.

    abstract::Cross-reactive carbohydrate determinants of plants are essentially a mixture of N-glycans containing beta1,2-xylose and core alpha1,3-fucose, the latter also found in insect glycoproteins. To determine the relative contributions of these two sugar residues to antibody binding, we prepared an array of glycomodified for...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh058

    authors: Bencúrová M,Hemmer W,Focke-Tejkl M,Wilson IB,Altmann F

    更新日期:2004-05-01 00:00:00

  • Ganglioside DSGb5, preferred ligand for Siglec-7, inhibits NK cell cytotoxicity against renal cell carcinoma cells.

    abstract::In renal cell carcinoma (RCC), the presence of higher gangliosides correlates with systematic metastasis. Disialosyl globopentaosylceramide (DSGb5) was identified previously as one of the major gangliosides from RCC tissues. Siglec-7 (sialic acid-binding Ig-like lectin-7), expressed on natural killer (NK) cells as an ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwq116

    authors: Kawasaki Y,Ito A,Withers DA,Taima T,Kakoi N,Saito S,Arai Y

    更新日期:2010-11-01 00:00:00

  • Advances in the use of biotinylated diaminopyridine (BAP) as a versatile fluorescent tag for oligosaccharides.

    abstract::We recently described a novel fluorescent compound, 2-amino,6-amidobiotinyl-pyridine (BAP), that allows the tagging of oligosaccharides, their fractionation by reversed-phase HPLC with picomole scale detection, and the formation of functional neoglycoprotein equivalents with (strept) avidin for the detection of recept...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/4.5.653

    authors: Toomre DK,Varki A

    更新日期:1994-10-01 00:00:00

  • Chondroitin sulfate N-acetylgalactosaminyltransferase-1 (CSGalNAcT-1) involved in chondroitin sulfate initiation: Impact of sulfation on activity and specificity.

    abstract::Glycosaminoglycan (GAG) assembly initiates through the formation of a linkage tetrasaccharide region serving as a primer for both chondroitin sulfate (CS) and heparan sulfate (HS) chain polymerization. A possible role for sulfation of the linkage structure and of the constitutive disaccharide unit of CS chains in the ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwr172

    authors: Gulberti S,Jacquinet JC,Chabel M,Ramalanjaona N,Magdalou J,Netter P,Coughtrie MW,Ouzzine M,Fournel-Gigleux S

    更新日期:2012-04-01 00:00:00

  • Antiganglioside antibodies and their pathophysiological effects on Guillain-Barré syndrome and related disorders--a review.

    abstract::Guillain-Barré syndrome (GBS) is an acute immune-mediated polyradiculoneuropathy which can cause acute quadriplegia. Infection with micro-organisms, including Campylobacter jejuni (C. jejuni), Haemophilus influenzae, and Cytomegalovirus (CMV), is recognized as a main triggering event for the disease. Lipooligosacchari...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/cwp027

    authors: Kaida K,Ariga T,Yu RK

    更新日期:2009-07-01 00:00:00

  • Crystal structures of a β-trefoil lectin from Entamoeba histolytica in monomeric and a novel disulfide bond-mediated dimeric forms.

    abstract::β-Trefoil lectins are galactose/N-acetyl galactosamine specific lectins, which are widely distributed across all kingdoms of life and are known to perform several important functions. However, there is no report available on the characterization of these lectins from protozoans. We have performed structural and biophy...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwaa001

    authors: Khan F,Kurre D,Suguna K

    更新日期:2020-07-20 00:00:00

  • Physiological and glycomic characterization of N-acetylglucosaminyltransferase-IVa and -IVb double deficient mice.

    abstract::N-Acetylglucosaminyltransferase-IV (GnT-IV) has two isoenzymes, GnT-IVa and GnT-IVb, which initiate the GlcNAcbeta1-4 branch synthesis on the Manalpha1-3 arm of the N-glycan core thereby increasing N-glycan branch complexity and conferring endogenous lectin binding epitopes. To elucidate the physiological significance...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwp200

    authors: Takamatsu S,Antonopoulos A,Ohtsubo K,Ditto D,Chiba Y,Le DT,Morris HR,Haslam SM,Dell A,Marth JD,Taniguchi N

    更新日期:2010-01-01 00:00:00

  • Carbo-loading in Coccidioides spp.: A quantitative analysis of CAZyme abundance and resulting glycan populations.

    abstract::Coccidioides spp. are important pneumonia-causing pathogens of the American southwest, but little is known about their glycobiology and how their glycosylations differ from other pneumonia-causing fungi. There is mounting preliminary evidence to suggest genus or even species-specific glycosylations in the fungal kingd...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwz092

    authors: Mitchell NM,Grys TE,Lake DF

    更新日期:2020-02-19 00:00:00

  • Significant structural change in both O- and N-linked carbohydrate moieties of the antigenic galactomannan from Aspergillus fumigatus grown under different culture conditions.

    abstract::Invasive aspergillosis is an important cause of morbidity and mortality in immunocompromised patients. Diagnosis of this infection frequently employs detection of the circulating galactomannan in the patient serum using enzyme immunoassay (EIA), a highly sensitive and specific system. Although there are many structura...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwu091

    authors: Kudoh A,Okawa Y,Shibata N

    更新日期:2015-01-01 00:00:00

  • Sialic acid content of tissue-specific gp96 and its potential role in modulating gp96-macrophage interactions.

    abstract::Cancer-derived heat shock protein gp96 induces a tumor-specific protective immune response primarily mediated by cytotoxic T lymphocytes (CTL) directed toward cancer-associated peptides associated with gp96. Both innate and adaptive immune responses have been demonstrated using a cell culture-based signaling mechanism...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwp096

    authors: Suriano R,Ghosh SK,Chaudhuri D,Mittelman A,Banerjee A,Tiwari RK

    更新日期:2009-12-01 00:00:00

  • Associations of genetic polymorphisms of Siglecs with human diseases.

    abstract::Genetic polymorphism studies in humans provide unique opportunities to understand human biology and the mechanisms of diseases. Correlations between polymorphisms in the genes encoding human Siglecs and various diseases have been reported. Leading examples, such as the CD33 polymorphism associated with late-onset Alzh...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/cwu043

    authors: Angata T

    更新日期:2014-09-01 00:00:00

  • Computational analysis of multivalency in lectins: structures of garlic lectin-oligosaccharide complexes and their aggregates.

    abstract::Multivalency in lectins is a phenomenon that has been discussed at considerable length. The structural basis for the role of multivalency in garlic lectin has been investigated here through computational studies. Biochemical studies have shown that the binding affinity of garlic lectin for high mannose oligosaccharide...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwg095

    authors: Ramachandraiah G,Chandra NR,Surolia A,Vijayan M

    更新日期:2003-11-01 00:00:00

  • Impact of human galectin-1 binding to saccharide ligands on dimer dissociation kinetics and structure.

    abstract::Endogenous lectins can control critical biological responses, including cell communication, signaling, angiogenesis and immunity by decoding glycan-containing information on a variety of cellular receptors and the extracellular matrix. Galectin-1 (Gal-1), a prototype member of the galectin family, displays only one ca...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cww052

    authors: Romero JM,Trujillo M,Estrin DA,Rabinovich GA,Di Lella S

    更新日期:2016-12-01 00:00:00

  • Differences in the apical and basolateral pathways for glycosaminoglycan biosynthesis in Madin-Darby canine kidney cells.

    abstract::Serglycin with a green fluorescent protein tag (SG-GFP) expressed in epithelial Madin-Darby canine kidney cells is secreted mainly (85%) into the apical medium, but the glycosaminoglycan (GAG) chains on the SG-GFP protein core secreted basolaterally (15%) carry most of the sulfate added during biosynthesis (Tveit et a...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwj075

    authors: Vuong TT,Prydz K,Tveit H

    更新日期:2006-04-01 00:00:00

  • 6-alkynyl fucose is a bioorthogonal analog for O-fucosylation of epidermal growth factor-like repeats and thrombospondin type-1 repeats by protein O-fucosyltransferases 1 and 2.

    abstract::Protein O-fucosyltransferase 1 (Pofut1) and protein O-fucosyltransferase 2 (Pofut2) add O-linked fucose at distinct consensus sequences in properly folded epidermal growth factor (EGF)-like repeats and thrombospondin type-1 (TSR) repeats, respectively. Glycan chain elongation past O-fucose can occur to yield a tetrasa...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cws140

    authors: Al-Shareffi E,Chaubard JL,Leonhard-Melief C,Wang SK,Wong CH,Haltiwanger RS

    更新日期:2013-02-01 00:00:00

  • Influenza virus infection of desialylated cells.

    abstract::Sialic acid has long been considered to be the sole receptor for influenza virus. The viral hemagglutinin (HA) is known to bind cell surface sialic acid, and sialic acids on viral glyco-proteins are cleaved by the viral neuraminidase (NA) to promote efficient release of progeny virus particles. However, NWS-Mvi, a mut...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/10.7.649

    authors: Stray SJ,Cummings RD,Air GM

    更新日期:2000-07-01 00:00:00

  • Prevention of the death of the rat axotomized hypoglossal nerve and promotion of its regeneration by bovine brain gangliosides.

    abstract::We have examined the time course of the neuronal death and regeneration of rat axotomized hypoglossal nerve with various conditions of the nerve resection, and established a useful system to measure neurotrophic activities of bioactive substances. In this system, neuronal death can be evaluated by counting surviving n...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/9.11.1247

    authors: Itoh M,Fukumoto S,Baba N,Kuga Y,Mizuno A,Furukawa K

    更新日期:1999-11-01 00:00:00

  • Differential analysis of site-specific glycans on plasma and cellular fibronectins: application of a hydrophilic affinity method for glycopeptide enrichment.

    abstract::Isolation of glycopeptides utilizing hydrogen bonding between glycopeptide glycans and a carbohydrate-gel matrix in the organic phase is useful for site-specific characterization of oligosaccharides of glycoproteins, when combined with mass spectrometry. In this study, recovery of glycopeptides was improved by includi...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwj019

    authors: Tajiri M,Yoshida S,Wada Y

    更新日期:2005-12-01 00:00:00

  • Structural characterization of novel oligosaccharides of cell-surface glycoproteins of Trypanosoma cruzi.

    abstract::Affinity-purified glycopeptides were prepared from Trypanosoma cruzi using the carbohydrate-specific monoclonal antibody WIC29.26. These glycopeptides contain rhamnose, fucose, xylose, and galactose, in the ratio 1:1:2:3. A series of oligosaccharides was released from the glycopeptides by mild acid hydrolysis, while, ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/6.8.869

    authors: Haynes PA,Ferguson MA,Cross GA

    更新日期:1996-12-01 00:00:00

  • Fucosyltransferase substrate specificity and the order of fucosylation in invertebrates.

    abstract::Core alpha1,6-fucosylation is a conserved feature of animal N-linked oligosaccharides being present in both invertebrates and vertebrates. To prove that the enzymatic basis for this modification is also evolutionarily conserved, cDNAs encoding the catalytic regions of the predicted Caenorhabditis elegans and Drosophil...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwi028

    authors: Paschinger K,Staudacher E,Stemmer U,Fabini G,Wilson IB

    更新日期:2005-05-01 00:00:00

  • A refined palate: bacterial consumption of host glycans in the gut.

    abstract::The human intestine houses a dense microbial ecosystem in which the struggle for nutrients creates a continual and dynamic selective force. Host-produced mucus glycans provide a ubiquitous source of carbon and energy for microbial species. Not surprisingly, many gut resident bacteria have become highly adapted to effi...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/cwt040

    authors: Marcobal A,Southwick AM,Earle KA,Sonnenburg JL

    更新日期:2013-09-01 00:00:00

  • Sialic acid glycoengineering using N-acetylmannosamine and sialic acid analogs.

    abstract::Sialic acids cap the glycans of cell surface glycoproteins and glycolipids. They are involved in a multitude of biological processes and aberrant sialic acid expression is associated with several pathologies. Sialic acids modulate the characteristics and functions of glycoproteins and regulate cell-cell as well as cel...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwz026

    authors: Moons SJ,Adema GJ,Derks MT,Boltje TJ,Büll C

    更新日期:2019-06-01 00:00:00

  • beta 1,4-Galactosyltransferase activity in B cells detected using a simple ELISA-based assay.

    abstract::Lymphocytic beta 1,4-galactosyltransferase (beta 1,4-GalTase, EC 2.4.1.38) activity was measured in B cells using a neoglyco-protein, N-acetylglucosamine-phenylisothiocyanate-bovine serum albumin (GlcNAc-pITC-BSA), as an acceptor substrate in a novel enzyme-linked immunosorbent assay (ELISA)-based method. This assay p...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/5.4.365

    authors: Keusch J,Lydyard PM,Isenberg DA,Delves PJ

    更新日期:1995-06-01 00:00:00

  • Cell surface components of Leishmania: identification of a novel parasite lectin?

    abstract::Protozoan parasites of the genus Leishmania have a glycoconjugate surface coat (the glycocalyx) that acts as the interface between the parasite and its external environment. The principal components of the glycocalyx, the lipophosphoglycans and the glycoinositolphospholipids, have a variety of functions that facilitat...

    journal_title:Glycobiology

    pub_type: 杂志文章,评审

    doi:10.1093/glycob/5.2.161

    authors: Smith DF,Rangarajan D

    更新日期:1995-03-01 00:00:00