Conformation of receptor adopted upon interaction with virus revealed by site-specific fluorescence quenchers and FRET analysis

Jürgen Wruss; Philipp D Pollheimer; Irene Meindl; Annett Reichel; Katrin Schulze; Wolfgang Schöfberger; Jacob Piehler; Robert Tampé; Dieter Blaas; Hermann J Gruber

doi:10.1021/ja807917t

Conformation of receptor adopted upon interaction with virus revealed by site-specific fluorescence quenchers and FRET analysis

J Am Chem Soc. 2009 Apr 22;131(15):5478-82. doi: 10.1021/ja807917t.

Authors

Jürgen Wruss¹, Philipp D Pollheimer, Irene Meindl, Annett Reichel, Katrin Schulze, Wolfgang Schöfberger, Jacob Piehler, Robert Tampé, Dieter Blaas, Hermann J Gruber

Affiliation

¹ Max F. Perutz Laboratories, Institute of Medicinal Biochemistry, Dr. Bohr Gasse 9/3, Medical University of Vienna, Vienna A-1030, Austria.

PMID: 19331321
DOI: 10.1021/ja807917t

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).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Binding Sites
Fluorescence
Fluorescence Resonance Energy Transfer
Genetic Engineering
Humans
Molecular Probe Techniques*
Protein Binding
Protein Conformation
Receptors, LDL / metabolism*
Rhinovirus / metabolism*

Substances

Receptors, LDL