Stimulation of human olfactory receptor 17-40 with odorants probed by surface plasmon resonance.
We report here the results of human olfactory receptor (OR) 17-40 stimulation with some odorants probed by means of the double-channel surface plasmon resonance platform NanoSPR-6. OR 17-40 tagged with N-terminal cmyc sequence was heterologously co-expressed with Galpha(olf) protein in yeast, and receptor-carrying nanosomes were prepared from yeast membrane fraction. Then, receptors were specifically captured via anti-cmyc antibody attached to the gold-coated substrate in orientated or random way. Measurement of odorants effects were carried out in the presence of GTP-gamma-S in differential mode in order to compensate bulk changes of refractive index. For the first time, biosensing efficiency of olfactory films was discussed in terms of their thickness and Galpha(olf) accessibility to GTP-gamma-S. Bell-shaped response profile with two maxima (near 1 nM and near 1 microM) was established for helional, which is documented as highly specific agonist of OR 17-40. Unrelated odorant heptanal used as control, did not evoke significant variations of differential signal.
Benilova
Irina
I
Laboratoire Ampère, Ecole Centrale de Lyon, 36 Avenue Guy de Collongue, 69134, Ecully Cedex, France. i_skry@yahoo.com
Chegel
Vladimir I
VI
Ushenin
Yuri V
YV
Vidic
Jasmina
J
Soldatkin
Alexey P
AP
Martelet
Claude
C
Pajot
Edith
E
Jaffrezic-Renault
Nicole
N
D015374:Biosensing Techniques / Q000379:methods*; D004563:Electrochemistry / Q000379:methods*; D006801:Humans; D015336:Molecular Probe Techniques; D009812:Odorants / Q000032:analysis*; D018035:Receptors, Odorant / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D018035:Receptors, Odorant