"Peptabody": a new type of high avidity binding protein.

Détails

ID Serval
serval:BIB_BE4DBBDBCE80
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
"Peptabody": a new type of high avidity binding protein.
Périodique
Proceedings of the National Academy of Sciences of the United States of America
Auteur⸱e⸱s
Terskikh A.V., Le Doussal J.M., Crameri R., Fisch I., Mach J.P., Kajava A.V.
ISSN
0027-8424
Statut éditorial
Publié
Date de publication
1997
Peer-reviewed
Oui
Volume
94
Numéro
5
Pages
1663-1668
Langue
anglais
Notes
Publication types: Journal Article Publication Status: ppublish
Résumé
A new type of high avidity binding molecule, termed "peptabody" was created by harnessing the effect of multivalent interaction. A short peptide ligand was fused via a semi-rigid hinge region with the coiled-coil assembly domain of the cartilage oligomeric matrix protein, resulting in a pentameric multivalent binding molecule. In the first peptabody (Pab-S) described here, a peptide (S) specific for the mouse B-cell lymphoma BCL1 surface Ig idiotype, was selected from a phage display library. A fusion gene was constructed encoding peptide S, followed by the 24 aa hinge region from camel IgG and a modified 55 aa cartilage oligomeric matrix protein pentamerization domain. The Pab-S fusion protein was expressed in Escherichia coli in a soluble form at high levels and purified in a single step by metal-affinity chromatography. Pab-S specifically bound the BCL1 surface idiotype with an avidity of about 1 nM, which corresponds to a 2 x 10(5)-fold increase compared with the affinity of the synthetic peptide S itself. Biochemical characterization showed that Pab-S is a stable homopentamer of about 85 kDa, with interchain disulfide bonds. Pab-S can be dissociated under denaturing and reducing conditions and reassociated as a pentamer with full-binding activity. This intrinsic feature provides an easy way to combine Pab molecules with two different peptide specificities, thus producing heteropentamers with bispecific and/or chelating properties.
Mots-clé
Animals, Binding, Competitive, Blotting, Western, Chromatography, Gel, Computer Simulation, Escherichia coli/genetics, Extracellular Matrix Proteins, Glycoproteins/chemistry, Glycoproteins/metabolism, Ligands, Lymphoma, B-Cell, Mice, Mice, Inbred BALB C, Models, Molecular, Peptides/chemistry, Peptides/genetics, Plasmids, Protein Binding, Protein Conformation, Receptors, Cell Surface/metabolism, Recombinant Fusion Proteins/chemistry, Recombinant Fusion Proteins/metabolism, Sequence Analysis, Signal Transduction, Tumor Cells, Cultured
Pubmed
Web of science
Open Access
Oui
Création de la notice
18/11/2009 11:23
Dernière modification de la notice
20/08/2019 15:32
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