Conformation of Receptor Adopted upon Interaction with Virus Revealed by Site-Specific Fluorescence Quenchers and FRETAnalysis
Sprache des Titels:
Englisch
Original Kurzfassung:
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-His6 were found to bind per
virus particle. In the present study, ring formation of V33333-His6 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-His6 was labeled with
fluorescein, and the C-terminus with a new quencher which was bound to the His6 tag with nanomolar
affinity (Kd ?10-8 M) in the presence of 2 ?M NiCl2.