Development of a competitive binding assay for the <i>Burkholderia cenocepacia</i> lectin BC2L-A and structure activity relationship of natural and synthetic inhibitors

Beshr, Ghamdan; Sommer, Roman; Hauck, Dirk; Siebert, David Chan Bodin; Hofmann, Anna; Imberty, Anne; Titz, Alexander

doi:10.1039/c5md00557d

Cited by 20 publications

(27 citation statements)

References 53 publications

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“…The crystals were soaked with 10 mM 2MeSeFuc (Sommer et al, 2016a) for 2 min and then placed in 80% saturated Li 2 SO 4 ($2.5 M) for cryoprotection prior mounting in a cryoloop and flash freezing in liquid nitrogen. For the All4MeMan (Beshr et al, 2016) complex, the crystals were directly transferred for 20 seconds in a solution containing 2.25 M Li 2 SO 4 and 10 mM ligand prior mounting and freezing. Data were collected at 100 K at ESRF, Grenoble, France on beamline BM30A-FIP for 2MeSeFuc/Lb-Tec2 complex and BM14 for All4MeMan/Lb-Tec2 complex using an ADSC Q315 CCD and a MARCCD detector respectively.…”

Section: Star+methodsmentioning

confidence: 99%

“…was also prepared (All4MeMan) (Beshr et al, 2016). Both synthetic ligands were located unambiguously in the electrondensity maps of the crystal structures in complex with Lb-Tec2.…”

Section: Recognition Of Methylated Glycans Bymentioning

confidence: 99%

“…Allyl 4-O-methyl-a-D-mannopyranoside (1) was synthesized following the protocol of Beshr et al (Beshr et al, 2016) and methyl 2-O-methyl-seleno-L-fucopyranoside was synthetized following the protocol of Sommer et al (Sommer et al, 2016a).…”

Section: Chemical Synthesismentioning

confidence: 99%

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Crystal Structures of Fungal Tectonin in Complex with O-Methylated Glycans Suggest Key Role in Innate Immune Defense

et al. 2018

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Innate immunity is the first line of defense against pathogens and predators. To initiate a response, it relies on the detection of invaders, where lectin-carbohydrate interactions play a major role. O-Methylated glycans were previously identified as non-self epitopes and conserved targets for defense effector proteins belonging to the tectonin superfamily. Here, we present two crystal structures of Tectonin 2 from the mushroom Laccaria bicolor in complex with methylated ligands, unraveling the molecular basis for this original specificity. Furthermore, they revealed the formation of a ball-shaped tetramer with 24 binding sites distributed at its surface, resembling a small virus capsid. Based on the crystal structures, a methylation recognition motif was identified and found in the sequence of many tectonins from bacteria to human. Our results support a key role of tectonins in innate defense based on a distinctive and conserved type of lectin-glycan interaction.

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Section: Star+methodsmentioning

confidence: 99%

“…was also prepared (All4MeMan) (Beshr et al, 2016). Both synthetic ligands were located unambiguously in the electrondensity maps of the crystal structures in complex with Lb-Tec2.…”

Section: Recognition Of Methylated Glycans Bymentioning

confidence: 99%

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Crystal Structures of Fungal Tectonin in Complex with O-Methylated Glycans Suggest Key Role in Innate Immune Defense

et al. 2018

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“…Besides fungal and animal tectonins that recognize O -methylated glycans in pathogens or parasites, numerous other lectins recognize such O -alkylated ligands, e.g., the pilus adhesin from Pseudomonas aeruginosa PAK [31] or PapG from Escherichia coli [32]. In contrast, methylation of lectin ligands can also prevent binding, as observed with O -methylated fucose and mannose for P. aeruginosa LecB or B. cenocepacia BC2L-A [33]. Thus, O -methylation of glycans can tune biological recognition events.…”

Section: Introductionmentioning

confidence: 99%

O-Alkylated heavy atom carbohydrate probes for protein X-ray crystallography: Studies towards the synthesis of methyl 2-O-methyl-L-selenofucopyranoside

Sommer¹,

Hauck²,

Varrot³

et al. 2016

Beilstein J. Org. Chem.

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Selenoglycosides are used as reactive glycosyl donors in the syntheses of oligosaccharides. In addition, such heavy atom analogs of natural glycosides are useful tools for structure determination of their lectin receptors using X-ray crystallography. Some lectins, e.g., members of the tectonin family, only bind to carbohydrate epitopes with O-alkylated ring hydroxy groups. In this context, we report the first synthesis of an O-methylated selenoglycoside, specifically methyl 2-O-methyl-L-selenofucopyranoside, a ligand of the lectin tectonin-2 from the mushroom Laccaria bicolor. The synthetic route required a strategic revision and further optimization due to the intrinsic lability of alkyl selenoglycosides, in particular for the labile fucose. Here, we describe a successful synthetic access to methyl 2-O-methyl-L-selenofucopyranoside in 9 linear steps and 26% overall yield starting from allyl L-fucopyranoside.

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“…In Pseudomonas aeruginosa, a ubiquitous opportunistic pathogen with a high current medical need for new therapeutics (14), the two lectins LecA and LecB are both toxins and adhesins with roles in biofilm formation and persistent infection (15)(16)(17). LecB (18,19)type proteins are relatively common, and several LecB homologs have also been previously characterized in other bacterial species such as Ralstonia solanaceum (20), Burkholderia cenocepacia (21)(22)(23), and Chromobacterium violaceum (24). The P. luminescens genome harbors several genes of predicted lectins that are homologs of LecB (25).…”

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confidence: 99%

Photorhabdus luminescens lectin A (PllA): A new probe for detecting α-galactoside–terminating glycoconjugates

Beshr

Sikandar

Jemiller

et al. 2017

Journal of Biological Chemistry

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Lectins play important roles in infections by pathogenic bacteria, for example, in host colonization, persistence, and biofilm formation. The Gram-negative entomopathogenic bacterium symbiotically lives in insect-infecting nematodes and kills the insect host upon invasion by the nematode. The genome harbors the gene, coding for a novel lectin that we named PllA. We analyzed the binding properties of purified PllA with a glycan array and a binding assay in solution. Both assays revealed a strict specificity of PllA for α-galactoside-terminating glycoconjugates. The crystal structures of apo PllA and complexes with three different ligands revealed the molecular basis for the strict specificity of this lectin. Furthermore, we found that a 90° twist in subunit orientation leads to a peculiar quaternary structure compared with that of its ortholog LecA from We also investigated the utility of PllA as a probe for detecting α-galactosides. The α-Gal epitope is present on wild-type pig cells and is the main reason for hyperacute organ rejection in pig to primate xenotransplantation. We noted that PllA specifically recognizes this epitope on the glycan array and demonstrated that PllA can be used as a fluorescent probe to detect this epitope on primary porcine cells In summary, our biochemical and structural analyses of the lectin PllA have disclosed the structural basis for PllA's high specificity for α-galactoside-containing ligands, and we show that PllA can be used to visualize the α-Gal epitope on porcine tissues.

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Development of a competitive binding assay for the Burkholderia cenocepacia lectin BC2L-A and structure activity relationship of natural and synthetic inhibitors

Abstract: Burkholderia cenocepacia is an opportunistic Gram-negative pathogen and especially hazardous for cystic fibrosis patients.

Cited by 20 publications

References 53 publications

Crystal Structures of Fungal Tectonin in Complex with O-Methylated Glycans Suggest Key Role in Innate Immune Defense

Crystal Structures of Fungal Tectonin in Complex with O-Methylated Glycans Suggest Key Role in Innate Immune Defense

O-Alkylated heavy atom carbohydrate probes for protein X-ray crystallography: Studies towards the synthesis of methyl 2-O-methyl-L-selenofucopyranoside

Photorhabdus luminescens lectin A (PllA): A new probe for detecting α-galactoside–terminating glycoconjugates

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