Distribution of Gal.alpha.1.fwdarw.3Gal.beta.1.fwdarw.4GlcNAc residues on secreted mammalian glycoproteins (thyroglobulin, fibrinogen, and immunoglobulin G) as measured by a sensitive solid-phase radioimmunoassay

Thall, Aron; Galili, Uri

doi:10.1021/bi00468a024

Cited by 133 publications

(68 citation statements)

References 44 publications

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“…As control reactions, we included transblots that were incubated with goat anti-human IgG alone (no anti-Gal). To assess the specificity ofthe anti-Gal binding to these lysates, we incubated transblots with Gal-serum as the blocking agent in place ofcasein, followed with biotinylated anti-Gal (20 ,g/ml) (25) and then avidin linked to alkaline phosphatase (as per Vector Laboratories). The blots were visualized as outlined above.…”

Section: Methodsmentioning

confidence: 99%

Human natural anti-Gal IgG regulates alternative complement pathway activation on bacterial surfaces.

Hamadeh¹,

Jarvis²,

Galili³

et al. 1992

J. Clin. Invest.

119

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One percent of circulating IgG in humans recognizes galactose al,3 galactose residues (anti-Gal) and is synthesized in response to stimulation by enteric bacteria. In this study, we found that the prevalence of binding of anti-Gal to blood isolates is significantly higher than its binding to normal stool isolates. When anti-Gal bound onto the lipopolysaccharide of a representative blood isolate, Serratia marcescens #21, it blocked its alternative complement pathway (ACP) lysis and made the organism serum resistant. In contrast, when anti-Gal bound to the capsular polysaccharide of a serum sensitive Serratia, #7, it increased ACP killing ofthis strain. The mechanism of blockade of ACP lysis by anti-Gal did not involve a decrease in the number ofC3 molecules deposited onto Serratia #21 or an inhibition of the binding of C3b to its LPS, nor did it change the iC3b and C3d degradation products of bound C3b or prevent membrane attack complex formation on this organism. Our findings suggest that the effect ofanti-Gal on immune lysis is dependent on the bacterial outer membrane structure to which it binds. We postulate that anti-Gal may play a role in the survival of selected Enterobacteriace in Gram-negative sepsis by blocking ACP-mediated lysis of such bacteria by the nonimmune host, and that this effect depends on where antiGal finds its epitope on the bacterial outer membrane. (J. Clin.

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Section: Methodsmentioning

confidence: 99%

Human natural anti-Gal IgG regulates alternative complement pathway activation on bacterial surfaces.

Hamadeh¹,

Jarvis²,

Galili³

et al. 1992

J. Clin. Invest.

119

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“…As a result, the mass loss of 527 u in a GONE sequencing step can only be explained by a Gal-GalGlcNAc sequence for mammalian glycans (see Figure 3a). The nonreducing end Gal in the Gal-Gal-GlcNAc sequence is most likely ␣ linked [56,57] as a terminal Gal(␤1-4)-Gal(␤1-4)-GlcNAc sequence in a N-linked glycan has only been found in fish [58] but not in mammals. Just as the mass loss of 365 u is diagnostic of a Gal-GlcNAc group, the mass loss of 527 u in GONE sequencing can be used as an indication of nonreducing end ␣Gal-Gal-GlcNAc for mammalian glycans.…”

Section: Fucosylated Bi-antennary With or Without ␣Galmentioning

confidence: 99%

Gas-phase oligosaccharide nonreducing end (GONE) sequencing and structural analysis by reversed phase hplc/mass spectrometry with polarity switching

Chen

Flynn

2009

J. Am. Soc. Mass Spectrom.

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Here we describe a technique to obtain all the N-linked oligosaccharide structures from a single reversed-phase (RP) HPLC run using on-line tandem MS in both positive and negative ion modes with polarity switching. Oligosaccharides labeled with 2-aminobenzamide (2AB) were used because they generated good ionization efficiency in both ion polarities. In the positive ion mode, protonated oligosaccharide ions lose sugar residues sequentially from the nonreducing end with each round of MS fragmentation, revealing the oligosaccharide sequence from greatly simplified tandem MS spectra. In the negative ion mode, diagnostic ions, including those from cross-ring cleavages, are readily observed in the MS 2 spectra of deprotonated oligosaccharide ions, providing detailed structural information, such as branch composition and linkage positions. Both positive and negative ion modes can be programmed into the same LC/MS experiment through polarity switching of the MS instrument. The gas-phase oligosaccharide nonreducing end (GONE) sequencing data, in combination with the diagnostic ions generated in negative ion tandem MS, allow both sequence and structural information to be obtained for all eluting species during a single RP-HPLC chromatographic run. This technique generates oligosaccharide analyses at high speed and sensitivity, and reveals structural features that can be difficult to obtain by traditional methods. . Proper structural determination is thus critical to the understanding of the biological role of carbohydrates. Although there are fewer monosaccharide building blocks in N-linked glycans than there are amino acid moieties in proteins, the combination of multiple branchings and linkages add greater complexity to the glycan structures. Such structural diversity also adds to the difficulty of structural characterization.Many analytical tools are now available for oligosaccharide characterization, such as high field NMR, liquid chromatography, capillary electrophoresis, and mass spectrometry. Although unambiguous oligosaccharide structural assignments can be made by NMR [2][3][4][5], this technique often requires large quantities of highly purified material. Typically, more than 50 mg of glycoprotein and extensive preparative fractionation are required to resolve oligosaccharide structures of relatively low abundance. A more common characterization approach utilizes sets of specific exoglycosidases, which cleave terminal monosaccharides from the nonreducing end, followed by chromatographic or MS analysis [6][7][8]. In this approach, the sequence of the oligosaccharide chain is revealed, either through sequential removal of monosaccharides from the chain upon successive enzyme treatment, or from the oligosaccharide ladder generated upon treatment with an array of enzymes. Linkage positional information and anomericity of the terminal monosaccharide can also be obtained, based on the specificity of the exoglycosidases used. Unfortunately, this classic sequencing technique is often labor intensive, time consuming, and...

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“…In humans and higher primates, the gene encoding alpha-1,3-galactosyltransferase is not functional and no α-Gal is synthesized [7]. Although IgG antibodies to α-Gal are widely expressed in humans [8], presumably in response to continuous exposure to the α-Gal epitope via gut microorganisms [9], IgE antibodies to α-Gal are not. An IgE antibody response to α-Gal was rst recognized when patients treated with the monoclonal antibody cetuximab experienced anaphylactic reactions upon the rst injection [10].…”

Section: Introductionmentioning

confidence: 99%

The red meat allergy syndrome in Sweden

Apostolović¹,

Tran²,

Starkhammar³

et al. 2016

Allergo J

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SummaryIn the last decade, a novel type of food allergy presenting with severe allergic reactions several hours a er consumption of red meat has been recognized.e allergic responses are due to IgE antibodies directed against the carbohydrate epitope galactose-α-1,3-galactose (α-Gal) found in mammalian meat.is review presents the red meat allergy syndrome in Sweden, discusses the features of the immune response to carbohydrates, and highlights the presence of heat stable α-Gal-containing proteins in meat.e number of diagnosed red meat allergy cases in Sweden has increased signi cantly over the past few years. All patients have been tick bitten. Our recent work has shown that α-Gal is present in the Euro pean tick Ixodes ricinus (I. ricinus), thus potentially explaining the strong association between anti-α-Gal IgE and tick bites, with development of red meat allergy as a secondary phenomenon. Further studies using immunoproteomics have identi ed novel α-Gal-containing meat proteins that bound IgE from red meat allergic patients. Four of these proteins were stable to thermal processing pointing to the fact that the allergenicity of red meat proteins is preserved in cooked meat. In keeping with the fact that the α-Gal epitope is structurally related to the blood group B antigen, a positive association with the B-negative blood groups among our red meat allergic patients was noted. A selective IgE reactivity to the pure carbohydrate moiety was observed when investigating the speci city of the α-Gal immune response. IgE from red meat allergic patients does not recognize the other major mammalian carbohydrate, N-glycolylneuraminic acid (Neu5Gc), also present in high amounts in red meat. Furthermore, neither common cross-reactive carbohydrate determinants (CCDs) from plants nor venoms are targets of the IgE response in these patients.Taken together, the α-Gal carbohydrate has shown to be a potentially clinically relevant allergen that should be taken into account in the diagnosis of food allergy. Many new ndings in the eld of red meat allergy have been obtained during the past years, but further e orts to understand the process of digestion, absorption, and delivery of α-Gal-containing molecules to the circulation are needed. e red meat allergy syndrome in Sweden. Allergo Cite this as

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Distribution of Gal.alpha.1.fwdarw.3Gal.beta.1.fwdarw.4GlcNAc residues on secreted mammalian glycoproteins (thyroglobulin, fibrinogen, and immunoglobulin G) as measured by a sensitive solid-phase radioimmunoassay

Cited by 133 publications

References 44 publications

Human natural anti-Gal IgG regulates alternative complement pathway activation on bacterial surfaces.

Human natural anti-Gal IgG regulates alternative complement pathway activation on bacterial surfaces.

Gas-phase oligosaccharide nonreducing end (GONE) sequencing and structural analysis by reversed phase hplc/mass spectrometry with polarity switching

The red meat allergy syndrome in Sweden

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