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

Lectins play important roles in infections by pathogenic bacteria, for example, in host colonization, persistence, and biofilm formation. The Gram-negative entomopathogenic bacterium Photorhabdus luminescens symbiotically lives in insect-infecting Heterorhabditis nematodes and kills the insect host...

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Veröffentlicht in:The Journal of biological chemistry 2017-12, Vol.292 (48), p.19935-19951
Hauptverfasser: Beshr, Ghamdan, Sikandar, Asfandyar, Jemiller, Eva-Maria, Klymiuk, Nikolai, Hauck, Dirk, Wagner, Stefanie, Wolf, Eckhard, Koehnke, Jesko, Titz, Alexander
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container_end_page 19951
container_issue 48
container_start_page 19935
container_title The Journal of biological chemistry
container_volume 292
creator Beshr, Ghamdan
Sikandar, Asfandyar
Jemiller, Eva-Maria
Klymiuk, Nikolai
Hauck, Dirk
Wagner, Stefanie
Wolf, Eckhard
Koehnke, Jesko
Titz, Alexander
description Lectins play important roles in infections by pathogenic bacteria, for example, in host colonization, persistence, and biofilm formation. The Gram-negative entomopathogenic bacterium Photorhabdus luminescens symbiotically lives in insect-infecting Heterorhabditis nematodes and kills the insect host upon invasion by the nematode. The P. luminescens genome harbors the gene plu2096, 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 Pseudomonas aeruginosa. 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 vitro. In summary, our biochemical and structural analyses of the P. luminescens 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.
doi_str_mv 10.1074/jbc.M117.812792
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subjects Amino Acid Sequence
Animals
carbohydrate
carbohydrate-binding protein
Galactosides - metabolism
glycobiology
Glycobiology and Extracellular Matrices
Glycoconjugates - metabolism
Hemagglutination Tests
lectin
Lectins - chemistry
Lectins - isolation & purification
Lectins - metabolism
Molecular Probes
Photorhabdus - metabolism
Protein Binding
Protein Conformation
protein structure
Sequence Homology, Amino Acid
structural biology
Swine
title Photorhabdus luminescens lectin A (PllA): A new probe for detecting α-galactoside–terminating glycoconjugates
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