Characterization of the five-fold Ca2+ binding site of satellite tobacco necrosis virus using Eu3+ luminescence spectroscopy: a marked size-selectivity among rare earth ions.
Satellite tobacco necrosis virus (STNV) is an icosahedral virus which contains three classes of Ca2+ binding site. One of these classes, a five-fold carbonyl site which is believed to exist in a Ca2+ channel, has been investigated using laser-induced Eu3+ luminescence spectroscopy. These twelve identical sites are rather rigid, as evidenced by the single narrow (full width at half-maximum is 6.5 cm-1) band observed at 579.58 nm in the 7F0----5D0 excitation spectrum of the Eu(3+)-STNV complex. Lifetimes of 270 microseconds in H2O and 1620 microseconds in D2O indicate that there are three water molecules bound to the Eu3+ at this site. Ligand field splitting of the 7F0----5D1 and 7F0----5D2 excitation spectra show that this site possesses fairly high symmetry (less than or equal to C5V). The Eu3+ complex of nitrilotriacetic acid was determined via titration to have a dissociation constant, Kd, of 20 +/- 2 nM; this value has been used in competition experiments to deduce that the virus site class binds Eu3+ with a Kd of 1.1 +/- 0.3 nM. This putative ion channel demonstrates remarkable size selectivity, with lanthanide affinities varying by more than one order of magnitude.
Burroughs
S E
SE
Department of Chemistry, Pennsylvania State University, University Park, 16802.
Eisenman
G
G
Horrocks
W D
WD
D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002413:Cations, Divalent; D005063:Europium; D008163:Luminescent Measurements; D010942:Plant Viruses / Q000378:metabolism*; D012526:Satellite Viruses / Q000378:metabolism*; D013057:Spectrum Analysis
D002413:Cations, Divalent; D005063:Europium; D002118:Calcium