IgE-binding properties and selectivity of peptide mimics of the FcvarepsilonRI binding site.
FcvarepsilonRIalpha found on the surface of mast cells and basophiles mediates allergic diseases, anaphylaxis and asthma through binding of IgE. Disrupting this interaction with anti-IgE mAbs has proven an efficient approach to control these diseases. The crystallographic structure of the complex formed between the IgE-Fc and FcvarepsilonRIalpha extracellular domain has shown that recognition is mediated by residues in the second Ig-like domain of the receptor (D2) and in the loop connecting the D1 and D2 domains. In an attempt to obtain specific IgE antagonists, we have designed and prepared a polypeptide named IgE-Trap that partially reproduces the IgE receptor-binding sites and binds with micromolar affinity to soluble IgE. The polypeptide contains loops C'-E [residues 129-134] and F-G [residues 151-161] from the D2 domain joined by a linker, and loop B-C [residues 110-113]. Peptide binding to IgE has been assessed by SPR analyses and the data fit with a biphasic model of interaction, in agreement with the two-site mechanism reported for the native receptor. The polypeptide binds to immobilized IgE in a dose-dependent manner with a K(D) estimated to be around 6muM, while it does not recognize IgG nor IgA. Polypeptide sub-domains involved in IgE binding have also been defined, showing that loop C'-E connected to loop B-C, but also the isolated loop B-C alone suffice to bind immunoglobulins E with high selectively though with reduced affinity compared to IgE-Trap. ELISA and cytometric assays on RBL2H3 cells demonstrate that the interacting peptides are able to displace the binding of IgE to receptor, confirming affinity and specificity of these ligands and suggesting a potential application as modulators of disorders associated with inappropriate IgE production.
Sandomenico
Annamaria
A
Istituto di Biostrutture e Bioimmagini, CNR, Napoli, Italy.
Monti
Simona M
SM
Marasco
Daniela
D
Dathan
Nina
N
Palumbo
Rosanna
R
Saviano
Michele
M
Ruvo
Menotti
M
D001665:Binding Sites; D002460:Cell Line; D006967:Hypersensitivity / Q000188:drug therapy / Q000378:metabolism; D007073:Immunoglobulin E / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017455:Receptors, IgE / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
C475664:FCER1A protein, human; D008024:Ligands; D010455:Peptides; D017455:Receptors, IgE; D007073:Immunoglobulin E