TAILIEUCHUNG - Báo cáo khoa học: Creating hybrid proteins by insertion of exogenous peptides into permissive sites of a class A b-lactamase

Fibronectins (FNs) are dimeric glycoproteins that adopt a globular con-formation when present in plasma and solution and an extended confor-mation in the extracellular matrix. Factor XII (FXII) is a zymogen of the proteolytically active FXIIa that plays a role in thrombus stabiliza-tion by enhancing clot formation and in inflammation by enhancing bradykinin formation. | ễFEBS Journal Creating hybrid proteins by insertion of exogenous peptides into permissive sites of a class A b-lactamase Nadia Ruth1 z Birgit Quinting1 f Jacques Mainil2 Bernard Hallet3 Jean-Marie Frere1 Kris Huygen4 and Moreno Galleni1 1 BiologicalMacromolecules and Laboratory of Enzymology Centre d Ingenierie des Proteines University of Liege Belgium 2 Laboratoire de Bacteriologie Departement des Maladies Infectieuses et Parasitaires University of Liege Belgium 3 Unite de Genetique Institut des Sciences de la Vie Universite Catholique de Louvain Louvain-la-Neuve Belgium 4 Mycobacteriallmmunology WIV-Pasteur Institute of Brussels Belgium Keywords class A b-lactamase TEM-1 hybrid protein insertion STa enterotoxin V3 loop Insertion of heterologous peptide sequences into a protein carrier may impose structural constraints that could help the peptide to adopt a proper fold. This concept could be the starting point for the development of a new Correspondence M. Galleni BiologicalMacromolecules CIP Sart-Tilman University of Liege B 4000 Liege Belgium Fax 32 4 366 33 64 Tel 32 4 366 35 49 E-mail mgalleni@ generation of safe subunit vaccines based on the expression of poorly immunogenic epitopes. In the present study we characterized the tolerance of the TEM-1 class A b-lactamase to the insertion of two different peptides the V3 loop of the gp120 protein of HIV and the thermostable STa enterotoxin produced by enterotoxic Escherichia coli. Insertion of the V3 loop of the HIV gp120 protein into the TEM-1 scaffold yielded insoluble and poorly produced proteins. By contrast four hybrid b-lactamases These authors contributed equally to this work carrying the STa peptide were efficiently produced and purified. Immunization of BALB c mice with these hybrid proteins produced high levels of TEM-1-specific antibodies together with significant levels of neutralizing Present address Division Immunologie Animale CER groupe Marloie Belgium antibodies against STa. Received 16 .

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