title: Conformation of receptor adopted upon interaction with virus revealed by site-specific fluorescence quenchers and FRET analysis.
abstract: Human rhinovirus serotype 2 (HRV2) specifically binds to very-low-density lipoprotein receptor (VLDLR). Among the eight extracellular repeats of VLDLR, the third module (V3) has the highest affinity for the virus, and 12 copies of the genetically engineered concatamer V33333-His(6) were found to bind per virus particle. In the present study, ring formation of V33333-His(6) about each of the 12 5-fold symmetry axes on HRV2 was demonstrated by fluorescence resonance energy transfer (FRET) between donor and acceptor on N- and C-terminus, respectively. In particular, the N-terminus of V33333-His(6) was labeled with fluorescein, and the C-terminus with a new quencher which was bound to the His(6) tag with nanomolar affinity (K(d) approximately 10(-8) M) in the presence of 2 microM NiCl(2).
authors.affiliation: Max F. Perutz Laboratories, Institute of Medicinal Biochemistry, Dr. Bohr Gasse 9/3, Medical University of Vienna, Vienna A-1030, Austria.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Wruss
authors.forename: Philipp D
authors.initials: PD
authors.lastname: Pollheimer
authors.forename: Irene
authors.initials: I
authors.lastname: Meindl
authors.forename: Annett
authors.initials: A
authors.lastname: Reichel
authors.forename: Katrin
authors.initials: K
authors.lastname: Schulze
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Schöfberger
authors.forename: Jacob
authors.initials: J
authors.lastname: Piehler
authors.forename: Robert
authors.initials: R
authors.lastname: Tampé
authors.forename: Dieter
authors.initials: D
authors.lastname: Blaas
authors.forename: Hermann J
authors.initials: HJ
authors.lastname: Gruber