Receptor phage

Scarselli, Elisa; Esposito, Gloria; Traboni, Cinzia

doi:10.1016/0014-5793(93)80226-k

Cited by 34 publications

(10 citation statements)

References 21 publications

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“…However, the role of the first domain is not clear in Fc⑀RI nor indeed in any Fc receptor. Analyses to date have variously indicated that domain one is necessary for optimal binding (1)(2)(3), that it has a possible role in direct interaction with IgE (4,5), and that it provides a supportive role in maintaining receptor integrity (1,2). The structural reasons for this are not apparent.…”

mentioning

confidence: 99%

Domain One of the High Affinity IgE Receptor, FcεRI, Regulates Binding to IgE through Its Interface with Domain Two

Rigby¹,

Epa²,

Mackay³

et al. 2000

Journal of Biological Chemistry

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The high affinity receptor for IgE, Fc⑀RI, binds IgE through the second Ig-like domain of the ␣ subunit. The role of the first Ig-like domain is not well understood, but it is required for optimal binding of IgE to Fc⑀RI, either through a minor contact interaction or in a supporting structural capacity. The results reported here demonstrate that domain one of Fc⑀RI plays a major structural role supporting the presentation of the ligand-binding site, by interactions generated within the interdomain interface. Analysis of a series of chimeric receptors and point mutants indicated that specific residues within the A strand of domain one are crucial to the maintenance of the interdomain interface, and IgE binding. Mutation of the Arg 15 and Phe 17 residues caused loss in ligand binding, and utilizing a homology model of Fc⑀RI-␣ based on the solved structure of Fc␥RIIa, it appears likely that this decrease is brought about by collapse of the interface and consequently the IgE-binding site. In addition discrepancies in results of previous studies using chimeric IgE receptors comprising Fc⑀RI␣ with either Fc␥RIIa or Fc␥RIIIA can be explained by the presence or absence of Arg 15 and its influence on the IgE-binding site. The data presented here suggest that the second domain of Fc⑀RI-␣ is the only domain involved in direct contact with the IgE ligand and that domain one has a structural function of great importance in maintaining the integrity of the interdomain interface and, through it, the ligand-binding site.The high affinity IgE receptor, Fc⑀RI, is a tetrameric complex composed of an IgE-binding ␣ subunit associated with a tetraspan ␤ subunit and homodimeric ␥ subunits and is a key player in IgE dependent effector mechanisms. The ␣ subunit, Fc⑀RI-␣, is the ligand-binding chain and is composed of two Ig-like domains. The role of the second domain has been clearly defined as containing the IgE-binding region. However, the role of the first domain is not clear in Fc⑀RI nor indeed in any Fc receptor. Analyses to date have variously indicated that domain one is necessary for optimal binding (1-3), that it has a possible role in direct interaction with IgE (4, 5), and that it provides a supportive role in maintaining receptor integrity (1, 2). The structural reasons for this are not apparent. Fc⑀RI, however, is related to Fc␥RIIa, and the recent description of the three dimensional structure of Fc␥RIIa (6), Fc␥RIIb (7), and Fc⑀RI-␣ (8) may provide a basis for the understanding of the roles of the individual domains in Fc⑀RI and other Fc receptors.In the crystal structure of Fc␥RIIa the extracellular domains are "bent" to form an acute angle (52°) between domains 1 and 2. In this orientation, the IgG-binding site of domain 2 points away from the cell in such a manner as to be accessible to ligand, and domain 1 is angled away from the binding site and down toward the cell membrane. The acute angle is dictated by interactions within the interdomain interface, and the structural studies indicate that domain 1 is likely to su...

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mentioning

confidence: 99%

Domain One of the High Affinity IgE Receptor, FcεRI, Regulates Binding to IgE through Its Interface with Domain Two

Rigby¹,

Epa²,

Mackay³

et al. 2000

Journal of Biological Chemistry

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“…Extensive efforts were hence attempted to produce Cε3 and D2 as isolated domains or as engineered variants (e.g., fusion proteins) [27, 28, 29, 30, 31]. Engineering started when the 3D structure of IgE Fc and FcεRIα D1D2 was still unknown.…”

Section: A Synergic Ensemblementioning

confidence: 99%

“…The protein reached the cell surface (probed by an mAb recognizing a short peptide of D2), yet IgE binding was undetectable and membrane D2 was not recognized by FcεRIα-specific mAbs with IgE inhibitory activity, suggesting that the IgE binding site was not folded properly [27]. In the same year, two independent groups reported the expression of D2 on the surface of M13 bacteriophage as a fusion to protein gIII [28, 29], followed very recently by a D2 mutagenesis and phage display screening study [33]. Phage gIII protein fusion appeared as a more effective strategy to display D2, even though D2 binding to IgE was considerably reduced compared to phage-displayed D1D2 [28, 29].…”

Section: A Synergic Ensemblementioning

confidence: 99%

“…In the same year, two independent groups reported the expression of D2 on the surface of M13 bacteriophage as a fusion to protein gIII [28, 29], followed very recently by a D2 mutagenesis and phage display screening study [33]. Phage gIII protein fusion appeared as a more effective strategy to display D2, even though D2 binding to IgE was considerably reduced compared to phage-displayed D1D2 [28, 29]. As occurred for Cε3, extensive expression and refolding attempts failed to generate a stable D2 in E.coli , except for a rapidly aggregating version expressed in the periplasm [Vangelista, unpubl.…”

Section: A Synergic Ensemblementioning

confidence: 99%

“…G In an IL-2 receptor fusion the resulting membrane D2 is apparently misfolded, although no data are available on its possible dimerization [27]. H Phage display gIII fusion of D2 is interesting but controversial, no data on its monomeric/oligomeric state are available [28, 29, 33]. I A GST-fusion of D2 appears to be refolded from E. coli inclusion bodies, yet no data on its conformational state are available [33].…”

Section: A Synergic Ensemblementioning

confidence: 99%

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Current Progress in the Understanding of IgE-FcεRI Interaction

Vangelista

2003

Int Arch Allergy Immunol

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The last decade has seen a wealth of studies aimed at the characterization of the binding between IgE and its high-affinity receptor, FcΕRI. IgE-FcΕRI complex formation is a major molecular event in atopic allergy. IgE-FcΕRI binding connects allergen recognition to cellular triggering, ultimately leading to disease manifestations. Consequently, pharmacological intervention at this site is of universal relevance for atopic allergy. Until recent years, the complexity of IgE-FcΕRI binding, together with the difficulty in obtaining fully functional recombinant IgE and FcΕRI derivatives, often led to confusion and difficulty in data interpretation. Major advances in the understanding of this intricate protein-protein interaction have now been accomplished. Most of the current knowledge on the IgE-FcΕRI recognition mode derives from long-lasting efforts in the field of structural biology. Protein engineering, high-throughput screening, immunological and biochemical studies also made relevant contributions in this domain. The data accumulated to date predict that IgE and FcΕRI use their modular architecture to approach each other in an asymmetric stepwise manner determining a 1:1 stoichiometry. This recognition appears to be enhanced by conformational changes occurring upon binding, leading to the well-known high-affinity. In conclusion, the vast amount of high-quality data available broadened our knowledge on the IgE-FcΕRI system; however, the fine structural details of the recognition process are still largely hypothetical. More studies are necessary to provide the experimental comprehensive picture required to carefully design efficient drugs acting at the IgE-FcΕRI interface.

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Phage‐Displayed Epitopes as Bioreceptors for Biosensors

Atias

Lobel

Virta

et al. 2007

Handbook of Biosensors and Biochips

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“Phage display” is a powerful technique that allows expression of peptides or proteins on the phage surface. Exploitation of phage display technology will lead to isolation and production of a broad range of binders with predefined specificities. Emergent technologies based on phage display will benefit diagnostics by producing molecules that are otherwise unobtainable through traditional approaches. Exploitation of the advantages of phage display systems together with the ongoing development in the biosensor field would lead to revolutionary diagnostic devices. This chapter will focus on phage biology, the advantage of phage display, and its applications as epitope bioreceptors in biosensor use.

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Receptor phage

Cited by 34 publications

References 21 publications

Domain One of the High Affinity IgE Receptor, FcεRI, Regulates Binding to IgE through Its Interface with Domain Two

Domain One of the High Affinity IgE Receptor, FcεRI, Regulates Binding to IgE through Its Interface with Domain Two

Current Progress in the Understanding of IgE-FcεRI Interaction

Phage‐Displayed Epitopes as Bioreceptors for Biosensors

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