Toward a Comprehensive Molecular Analysis of Olfactory Transduction

Lancet, Doron; Chen, Zehava; Ciobotariu, Adina; Eckstein, Fritz; Khen, Miriam; Heldman, Judith; Ophir, Dov; Shafir, Iris; Pace, Umberto

doi:10.1111/j.1749-6632.1987.tb43462.x

Cited by 10 publications

(1 citation statement)

References 19 publications

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“…These receptors are predicted to contain seven transmembrane segments, consistent with the view that odorant signals are transduced via G-protein-coupled cascades in olfactory sensory neurons (Reed et al, 1992;Breer et al, 1994). Different receptor types are presumed to be tuned to odorous compounds with distinct chemical determinants, and odor coding may be accomplished by expression of one or a few receptor types in individual chemosensory neurons (Lancet et al, 1987;Shepherd, 1994;Mori & Yoshihara, 1995). Soluble ligand-specific odorant binding proteins have also been implicated in the decoding of odor and in signal transduction (Pelosi, 1994;Prestwich et al, 1995).…”

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confidence: 65%

Expression of an Olfactory Receptor inEscherichiacoli: Purification, Reconstitution, and Ligand Binding

et al. 1996

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An olfactory receptor has been expressed in bacterial cells as a fusion protein with glutathione S-transferase (GST). Overexpression of receptor protein yielding about 10% of the cell protein was achieved with mutants lacking the N-terminus and the first transmembrane region or with mutants carrying three positively charged residues in the first intracellular loop. The overexpressed fusion protein accumulated in inclusion bodies and could be solubilized in detergent. It was purified by metal chelation chromatography based on a C-terminal 6-histidine tag, and the GST portion was removed after proteolytic cleavage. The purified receptor was reconstituted into lipid vesicles and specific binding of odor ligands was shown by photoaffinity labeling and tryptophan fluorescence measurements. Thus, for the first time, an odorant receptor/ligand pair becomes available in large amounts for biophysical and screening studies.

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confidence: 65%