Differential Binding Properties of GalNAc and/or Gal Specific Lectins

Wu, Albert M.; Sugii, Shunji

doi:10.1007/978-1-4613-1663-3_9

Cited by 55 publications

(54 citation statements)

References 111 publications

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“…GP900 has previously been shown to be N glycosylated (26). The pattern of lectin binding to GP900 was consistent with the presence of internal or terminal ␣GalNAc residues (40). ␣-N-Acetylgalactosaminidase treatment had no discernible effect on 4E9 reactivity with GP900, suggesting that the ␣GalNAc residues may be internal or that if they are terminal, as in the case of gp40, a small shift in molecular weight may not be resolved due to the very high M r of the protein.…”

Section: Discussionmentioning

confidence: 53%

“…The pattern of reactivity of ␣GalNAc-specific lectins with gp40 is suggestive of the presence of the Tn determinant, which consists of terminal ␣GalNAc residues linked to Ser/Thr residues of the protein backbone (40). The presence of terminal ␣GalNAc residues in gp40 was further confirmed by the decrease in reactivity and shift in M r caused by ␣-N-acetylgalactosaminidase treatment.…”

Section: Discussionmentioning

confidence: 83%

“…These determinants are commonly present in core structures of the carbohydrate side chains of mucins. They are usually cryptic in O-linked oligosaccharides of mucins in normal tissue, but they may be terminally exposed in mucins on tumor cells (40). In addition to GalNAc, N-acetylglucosamine (GlcNAc) and galactose (Gal) O-glycans of human intestinal and other mucins contain peripheral sugars, including sialic acid, which mask the core structures.…”

Section: Discussionmentioning

confidence: 99%

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Mediation ofCryptosporidium parvumInfection In Vitro by Mucin-Like Glycoproteins Defined by a Neutralizing Monoclonal Antibody

et al. 2000

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The protozoan parasite Cryptosporidium parvum is a significant cause of diarrheal disease worldwide. Attachment to and invasion of host intestinal epithelial cells by C. parvum sporozoites are crucial steps in the pathogenesis of cryptosporidiosis. The molecular basis of these initial interactions is unknown. In order to identify putative C. parvum adhesion-and invasion-specific proteins, we raised monoclonal antibodies (MAbs) to sporozoites and evaluated them for inhibition of attachment and invasion in vitro. Using this approach, we identified two glycoproteins recognized by 4E9, a MAb which neutralized C. parvum infection and inhibited sporozoite attachment to intestinal epithelial cells in vitro. 4E9 recognized a 40-kDa glycoprotein named gp40 and a second, >220-kDa protein which was identified as GP900, a previously described mucin-like glycoprotein. Glycoproteins recognized by 4E9 are localized to the surface and apical region of invasive stages and are shed in trails from the parasite during gliding motility. The epitope recognized by 4E9 contains ␣-N-acetylgalactosamine residues, which are present in a mucin-type O-glycosidic linkage. Lectins specific for these glycans bind to the surface and apical region of sporozoites and block attachment to host cells. The surface and apical localization of these glycoproteins and the neutralizing effect of the MAb and ␣-N-acetylgalactosaminespecific lectins strongly implicate these proteins and their glycotopes as playing a role in C. parvum-host cell interactions.Cryptosporidium parvum, an intestinal Apicomplexan parasite, is a significant cause of diarrheal disease worldwide (15,17). In immunocompetent individuals, the disease is usually self-limiting, but it may be chronic and life threatening in immunocompromised patients such as those with AIDS. Recently, the parasite has gained notoriety as the causative agent of numerous outbreaks of waterborne diarrheal disease. There is currently no effective specific therapy approved for disease caused by this parasite.Infection is initiated by ingestion of oocysts, which undergo excystation to release sporozoites. Attachment of sporozoites to epithelial cells and subsequent invasion of the host cell membrane are crucial primary steps in the pathogenesis of cryptosporidiosis. The ultrastructural aspects of attachment and invasion have been characterized in detail (10,24,33,34). Sporozoites attach to host cells by their anterior pole. Attachment is followed by invagination of the host cell plasma membrane, which extends along the surface of the sporozoite and eventually completely surrounds it, leading to formation of a parasitophorus vacuole where the parasite undergoes further development in a unique intracellular but extracytoplasmic location.Using in vitro models of sporozoite attachment to epithelial cells, we previously showed that attachment was dose and time dependent and was influenced by pH, divalent cations, and the degree of differentiation of host cells (20,22). Further, attachment could be inhibited by polyclo...

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Section: Discussionmentioning

confidence: 53%

Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

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Mediation ofCryptosporidium parvumInfection In Vitro by Mucin-Like Glycoproteins Defined by a Neutralizing Monoclonal Antibody

et al. 2000

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“…In the present communication, the binding properties of the Sd(a-) THGP and its asialo product with a panel of lectins exhibiting a broad range of carbohydrate specificities were characterized by quantitative precipitin (QPA) and precipitin inhibition (QPIA) assays. During the past two decades, this system has been successfully used as a valuable tool to characterize the saccharide affinity of lectins [6,7,[27][28][29][30][31][32][33][34][35], as such studies can provide insight into the specificities and size parameters of the lectin-glycan interactions. Among 20 lectins (Table 1) tested by QPA, both native and asialo Sd(a-) THGP reacted best with APA and RCA1 and completely precipitated the lectin added (Fig.…”

Section: Resultsmentioning

confidence: 99%

Interaction of a human blood group Sd(a−) Tamm‐Horsfall glycoprotein with applied lectins

Watkins

Chen

et al. 1996

FEBS Letters

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Unlike the human blood group Sd(a+) Tamm-Horsfall glycoprotein (THGP), the Sd(a-) one lacks terminal GalNAc[~I--, residues at the nonreducing ends. The binding properties of this glycoprotein and its asialo product with lectins were characterized by quantitative precipitin (QPA) and precipitin inhibition assays. Among 20 lectins tested by QPA, both native and asialo Sd(a-) THGP reacted best with Abrus precatorius and Ricinus communis and completely precipitated the lectin added. They also precipitated well Wistariafloribunda (WFA), Glycine max (SBA), Bauhinia purpurea alba, abrin-a and ricin, all of which recognize the Gall31--, 4GIcNAc[~I--, sequence, although at different strength. The lectin-glyean interactions were inhibited by GaI[~I~4GIcNAc and Gal[~l --* 4Glc. When the precipitability of Sd(a-) THGP was compared with that of the Sd(a+) phenotype, the native Sd(a-) THGP exhibited a 40% lesser affinity for WFA, SBA, WGA and mistletoe lectin-I (ML-I). Mapping the precipitation and inhibition profiles of the present study and the results of THGP Sd(a+), it is concluded that Sd(a-) THGP showed a strongly diminished affinity for GalNAcl31-~ active lectins (SBA and WFA) than the Sd(a+) phenotype.

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“…Since the zoonotic species, C. parvum, is the most easily propagated in calves, research is done primarily with these parasites, despite the observation that C. hominis is the species responsible for most human infections [2,18]. Adding to the difficulties, the mucin-like glycoproteins that Cryptosporidium may rely on for attachment and invasion display an unusual "primitive " pattern of O-glycosylation with terminal α-N-acetyl-D-galactosamine (GalNAc) and Galβ1-3GalNAc (T and Tn antigen) determinants [19] which are normally cryptic on mammalian cells due to sialyation [20]. An ideal expression system for cryptosporidial proteins would have to produce similar glycosylation patterns.…”

Section: Introductionmentioning

confidence: 99%

Stable expression of Cryptosporidium parvum glycoprotein gp40/15 in Toxoplasma gondii

O’Connor

Wanyiri

Wójczyk

et al. 2007

Molecular and Biochemical Parasitology

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Cryptosporidium is a cause of diarrheal disease worldwide. Parasite glycoproteins involved in invasion of Cryptosporidium into host cells have been investigated as possible targets for effective interventions against this parasite. One of these, Cpgp40/15, is expressed as a precursor protein that is cleaved by a parasite-derived furin-like protease activity into gp15, a glycophosphatidyl inositol anchored surface protein, and gp40, that associates with gp15 and binds to host cells. Investigation of the functions of these glycoproteins requires an expression system that can produce similar glycosylation patterns to the native antigens. Previous work demonstrated that Cpgp40/15 transiently expressed in T. gondii was appropriately localized and glycosylated. In this study, T. gondii stable transfectants expressing gp40/15, gp15, gp40 and hemagglutinin (HA) tagged gp40 were generated. T. gondii recombinant gp40HA and gp40/15 (recTggp40HA and recTggp40/15) were isolated from infected cells by HA affinity chromatography and Helix pomatia lectin affinity chromatography, respectively. Mass spectrometry confirmed that recTggp40-HA and native Cpgp40 were similarly glycosylated. Like native Cpgp40/15, recTggp40/15 could be cleaved into the gp40 and gp15 products by human furin or by a furin-like protease activity in T. gondii tachyzoite lysates. However, processing was inefficient in intact tachyzoites. Unlike the N-terminus of native Cpgp40/15, which appears to be processed following signal peptide cleavage, the N-terminus of recTggp40/15 began at the predicted signal sequence cleavage site, 11 amino acids upstream of the N-terminus of native Cpgp40. The ability to express and isolate appropriately glycosylated Cryptosporidium glycoproteins will enable further investigations into host-parasite interactions of this important pathogen.

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Differential Binding Properties of GalNAc and/or Gal Specific Lectins

Cited by 55 publications

References 111 publications

Mediation ofCryptosporidium parvumInfection In Vitro by Mucin-Like Glycoproteins Defined by a Neutralizing Monoclonal Antibody

Mediation ofCryptosporidium parvumInfection In Vitro by Mucin-Like Glycoproteins Defined by a Neutralizing Monoclonal Antibody

Interaction of a human blood group Sd(a−) Tamm‐Horsfall glycoprotein with applied lectins

Stable expression of Cryptosporidium parvum glycoprotein gp40/15 in Toxoplasma gondii

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