Engineering Regulable Escherichia coli β-Galactosidases as Biosensors for Anti-HIV Antibody Detection in Human Sera

verfasst von
Neus Ferrer-Miralles, Jordi X. Feliu, Stéphane Vandevuer, Annette Müller, Joaquin Cabrera-Crespo, Isabelle Ortmans, Frank Hoffmann, Daniel Cazorla, Ursula Rinas, Martine Prévost, Antonio Villaverde
Abstract

The activity of engineered, peptide-displaying enzymes is modulated by binding to specific anti-peptide antibodies. This new concept of a quantitative antibody detection system allows test kits to be set up for fast diagnosis of infectious diseases. To develop a quick and homogeneous assay for the detection of human immunodeficiency virus (HIV) infection, we have explored two acceptor sites of the bacterial Escherichia coli β-galactosidase for the accommodation of HIV antigenic peptides. Two overlapping epitopes (namely P1 and P2) from the gp41 envelope glycoprotein, contained in different sized peptides, were inserted in the vicinity of the enzyme active site to generate a set of hybrid, enzymatically active β-galactosidases. Regulable enzymes of different responsiveness to monoclonal antibody binding were generated with both acceptor sites tested. These biosensors were also sensitive to immune sera from HIV-infected patients. Modeling data provide insight into the structural modifications in the vicinity of the active site induced by peptide insertion that strongly affect the responsiveness of the engineered proteins through different parameters of their catalytic properties.

Externe Organisation(en)
Universidad Autónoma de Barcelona (UAB)
Universität Pompeu Fabra (UPF)
Université libre de Bruxelles (ULB)
MicroCoat Biotechnologie GmbH
Helmholtz-Zentrum für Infektionsforschung GmbH (HZI)
Martin-Luther-Universität Halle-Wittenberg
Typ
Artikel
Journal
Journal of Biological Chemistry
Band
276
Seiten
40087-40095
Anzahl der Seiten
9
ISSN
0021-9258
Publikationsdatum
26.10.2001
Publikationsstatus
Veröffentlicht
Peer-reviewed
Ja
ASJC Scopus Sachgebiete
Biochemie, Molekularbiologie, Zellbiologie
Ziele für nachhaltige Entwicklung
SDG 3 – Gute Gesundheit und Wohlergehen
Elektronische Version(en)
https://doi.org/10.1074/jbc.M104704200 (Zugang: Geschlossen)