Interference of peptides and specific antibodies with the function of the Actinobacillus pleuropneumoniae transferrin-binding protein

Strutzberg, K; Franz, B; Gerlach, Gerald-F.

doi:10.1128/iai.65.12.5346-5348.1997

Cited by 3 publications

(1 citation statement)

References 17 publications

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“…Transposon mutagenesis of the tbpB gene revealed that TbpB-binding activity for pTf was retained in a fusion protein containing the first 289 amino acids of the mature protein and a series of randomly generated mutants with reduced binding activity primarily mapped to those regions (Strutzberg et al, 1995). A synthetic peptide library representing TbpB from A. pleuropneumoniae was probed with labeled pTf to reveal peptide regions with Tf-binding activity (Strutzberg et al, 1997). Similar approaches have been used to probe the tight interactions between TbpB and Tf in several human pathogens (Renauld-Monge ´nie et al, 1997;Retzer et al, 1999;Sims and Schryvers, 2003), but a clear understanding and appreciation of this interaction has been elusive.…”

Section: Introductionmentioning

confidence: 99%

Insights into the Bacterial Transferrin Receptor: The Structure of Transferrin-Binding Protein B from Actinobacillus pleuropneumoniae

Moraes

Strynadka

et al. 2009

Molecular Cell

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Pathogenic bacteria from the Neisseriaceae and Pasteurellacea families acquire iron directly from the host iron-binding glycoprotein, transferrin (Tf), in a process mediated by surface receptor proteins that directly bind host Tf, extract the iron, and transport it across the outer membrane. The bacterial Tf receptor is comprised of a surface exposed lipoprotein, Tf-binding protein B (TbpB), and an integral outer-membrane protein, Tf-binding protein A (TbpA), both of which are essential for survival in the host. In this study, we report the 1.98 A resolution structure of TbpB from the porcine pathogen Actinobacillus pleuropneumoniae, providing insights into the mechanism of Tf binding and the role of TbpB. A model for the complex of TbpB bound to Tf is proposed. Mutation of a single surface-exposed Phe residue on TbpB within the predicted interface completely abolishes binding to Tf, suggesting that the TbpB N lobe comprises the sole high-affinity binding region for Tf.

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Section: Introductionmentioning

confidence: 99%

Insights into the Bacterial Transferrin Receptor: The Structure of Transferrin-Binding Protein B from Actinobacillus pleuropneumoniae

Moraes

Strynadka

et al. 2009

Molecular Cell

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SPOT Synthesis — Scope of Applications

Frank

Schneider‐Mergener

2002

Peptide Arrays on Membrane Supports

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Specific Ligand Binding Attributable to Individual Epitopes of Gonococcal Transferrin Binding Protein A

Masri

Cornelissen

2002

Infect Immun

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The gonococcal transferrin receptor complex comprises two iron-regulated proteins, TbpA and TbpB. TbpA is essential for transferrin-iron uptake and is a TonB-dependent integral outer membrane protein. TbpB is thought to increase the efficiency of iron uptake from transferrin and is lipid modified and surface exposed. To evaluate the structure-function relationships in one of the components of the receptor, TbpA, we created constructs that fused individual putative loops of TbpA with amino-terminal affinity tags. The recombinant proteins were then overexpressed in Escherichia coli, and the fusions were recovered predominately from inclusion bodies. Inclusion body proteins were solubilized, and the epitope fusions were renatured by slow dialysis. To assess transferrin binding capabilities, the constructs were tested in a solid-phase dot blot assay followed by confirmatory quantitative chemiluminescent enzyme-linked immunosorbent assays. The constructs with only loop 5 and with loops 4 and 5 demonstrated dose-dependent specific ligand binding in spite of being out of the context of the intact receptor. The immunogenicities of individual TbpA-specific epitopes were investigated by generating rabbit polyclonal antisera against the fusion proteins. Most of the fusion proteins were immunogenic under these conditions, and the resulting sera recognized full-length TbpA in immunoblots. These results suggest that individual epitopes of TbpA are both immunogenic and functional with respect to ligand binding capabilities, and the vaccine implications of these findings are discussed.

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Interference of peptides and specific antibodies with the function of the Actinobacillus pleuropneumoniae transferrin-binding protein

Cited by 3 publications

References 17 publications

Insights into the Bacterial Transferrin Receptor: The Structure of Transferrin-Binding Protein B from Actinobacillus pleuropneumoniae

Insights into the Bacterial Transferrin Receptor: The Structure of Transferrin-Binding Protein B from Actinobacillus pleuropneumoniae

SPOT Synthesis — Scope of Applications

Specific Ligand Binding Attributable to Individual Epitopes of Gonococcal Transferrin Binding Protein A

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