Array-assisted Characterization of a Fucosyltransferase Required for the Biosynthesis of Complex Core Modifications of Nematode N-Glycans

Yan, Shi; Serna, Sonia; Reichardt, Niels‐Christian; Paschinger, Katharina; Wilson, Iain B. H.

doi:10.1074/jbc.m113.479147

Cited by 38 publications

(36 citation statements)

References 52 publications

(58 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2A). C. elegans fut-8 (ok2558) as well as the double mutants fut-6(ok475)fut-1(ok892) and fut-6(ok475);fut-8(ok2558), lacking fucose residues linked to the GlcNAc units of the N-glycan core (22)(23)(24), were equally susceptible toward Lb-Tec2 as the wild-type strain. We therefore concluded that α1,3-or α1,6-linked core fucose was not required for Lb-Tec2-induced nematotoxicity.…”

Section: Resultsmentioning

confidence: 99%

Methylated glycans as conserved targets of animal and fungal innate defense

Wohlschlager

Butschi

Grassi

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Effector proteins of innate immune systems recognize specific nonself epitopes. Tectonins are a family of β-propeller lectins conserved from bacteria to mammals that have been shown to bind bacterial lipopolysaccharide (LPS). We present experimental evidence that two Tectonins of fungal and animal origin have a specificity for O-methylated glycans. We show that Tectonin 2 of the mushroom Laccaria bicolor (Lb-Tec2) agglutinates Gram-negative bacteria and exerts toxicity toward the model nematode Caenorhabditis elegans, suggesting a role in fungal defense against bacteria and nematodes. Biochemical and genetic analysis of these interactions revealed that both bacterial agglutination and nematotoxicity of Lb-Tec2 depend on the recognition of methylated glycans, namely O-methylated mannose and fucose residues, as part of bacterial LPS and nematode cell-surface glycans. In addition, a C. elegans gene, termed samt-1, coding for a candidate membrane transport protein for the presumptive donor substrate of glycan methylation, S-adenosyl-methionine, from the cytoplasm to the Golgi was identified. Intriguingly, limulus lectin L6, a structurally related antibacterial protein of the Japanese horseshoe crab Tachypleus tridentatus, showed properties identical to the mushroom lectin. These results suggest that O-methylated glycans constitute a conserved target of the fungal and animal innate immune system. The broad phylogenetic distribution of O-methylated glycans increases the spectrum of potential antagonists recognized by Tectonins, rendering this conserved protein family a universal defense armor.

show abstract

Section: Resultsmentioning

confidence: 99%

Methylated glycans as conserved targets of animal and fungal innate defense

Wohlschlager

Butschi

Grassi

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…The six glycans G1-G6 were immobilized in different subarrays on NHS activated glass slides and individual wells incubated with bovine milk β-1,4 galactosyltransferase 2.4.1.22 (GalT), C. elegans β-1,4 Nacetylgalactosaminyltransferase (GalNAcT), Arabidopsis thaliana core type α-1,3 fucosyltransferase (AtFucTA), C. elegans core type α-1,3 fucosyltransferase (CeFUT1), C. elegans α-1,3 fucosyltransferase (CeFUT6) and C. elegans core type α-1,6 fucosyltransferase (CeFUT8). [22][23][24][25][26] The presence of enzymatically extended structures was verified by incubating wells with the fluorescently tagged lectins, Ricinus communis (RCA-I), Wisteria floribunda (WFL) and Aleuria aurantia (AAL-555) that specifically recognize β-galactose, N-acetylgalactosamine and α-fucose, respectively. 27 With the exception of CeFUT6, enzymatic elongations of xylosylated glycans were possible as long as other previously reported basic structural requirements were fulfilled.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and Microarray-Assisted Binding Studies of Core Xylose and Fucose Containing N-Glycans

et al. 2015

Self Cite

View full text Add to dashboard Cite

The synthesis of a collection of 33 xylosylated and core-fucosylated N-glycans found only in nonmammalian organisms such as plants and parasitic helminths has been achieved by employing a highly convergent chemo-enzymatic approach. The influence of these core modifications on the interaction with plant lectins, with the human lectin DC-SIGN (Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Nonintegrin), and with serum antibodies from schistosome-infected individuals was studied. Core xylosylation markedly reduced or completely abolished binding to several mannose-binding plant lectins and to DC-SIGN, a C-type lectin receptor present on antigen presenting cells. Employing the synthetic collection of core-fucosylated and core-xylosylated N-glycans in the context of a larger glycan array including structures lacking these core modifications, we were able to dissect core xylose and core fucose specific antiglycan antibody responses in S. mansoni infection sera, and we observed clear and immunologically relevant differences between children and adult groups infected with this parasite. The work presented here suggests that, quite similar to bisecting N-acetylglucosamine, core xylose distorts the conformation of the unsubstituted glycan, with important implications for the immunogenicity and protein binding properties of complex N-glycans.

show abstract

“…However, up to four fucose residues have been detected on C. elegans N-glycans and the exact nature of the linkage of the fourth fucose has remained obscure (3,12-14). Combined with the latest knowledge regarding the specificity of C. elegans core fucosyltransferases (13,15,16) as well as the exact structures of N-glycans from C. elegans and other nematodes (17), we concluded that some models for the tri- and tetrafucosylated N-glycans were incorrect. By preparing a triple mutant unable to core fucosylate its N-glycans, we generated a C. elegans strain containing maximally one fucose residue on the N-linked oligosaccharides.…”

Section: Introductionmentioning

confidence: 98%

Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans

Yan

Brecker

Jin

et al. 2015

Molecular & Cellular Proteomics

Self Cite

View full text Add to dashboard Cite

The N-glycosylation of the model nematode Caenorhabiditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that ‘simple’ organisms possess also a rather simple glycomic capacity. In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. However, although the linkage of three fucose residues to the N,N′-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. Within this glycomic sub-pool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a ‘bisecting’ β-galactose which is substoichiometrically modified with fucose or methylfucose. In addition, the α1,3-mannose can also be α-galactosylated. Not only do these residues constitute novel modifications of N-glycans, but these features may well be targets of nematoxic proteins produced as defence molecules by various fungal species.

show abstract

Array-assisted Characterization of a Fucosyltransferase Required for the Biosynthesis of Complex Core Modifications of Nematode N-Glycans

Cited by 38 publications

References 52 publications

Methylated glycans as conserved targets of animal and fungal innate defense

Methylated glycans as conserved targets of animal and fungal innate defense

Synthesis and Microarray-Assisted Binding Studies of Core Xylose and Fucose Containing N-Glycans

Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans

Contact Info

Product

Resources

About