Abstract
A crucial bottleneck in membrane protein studies, particularly G-protein coupled receptors, is the notorious difficulty of finding an optimal detergent that can solubilize them and maintain their stability and function. Here we report rapid production of 12 unique mammalian olfactory receptors using short designer lipid-like peptides as detergents. The peptides were able to solubilize and stabilize each receptor. Circular dichroism showed that the purified olfactory receptors had alpha-helical secondary structures. Microscale thermophoresis suggested that the receptors were functional and bound their odorants. Blot intensity measurements indicated that milligram quantities of each olfactory receptor could be produced with at least one peptide detergent. The peptide detergents' capability was comparable to that of the detergent Brij-35. The ability of 10 peptide detergents to functionally solubilize 12 olfactory receptors demonstrates their usefulness as a new class of detergents for olfactory receptors, and possibly other G-protein coupled receptors and membrane proteins.
Highlights
Olfactory receptors are arguably the most sensitive detectors: they distinguish between thousands of odorants down to parts per billion or trillion
Systematic screens were performed to assess the ability of peptide detergents to produce and solubilize 12 olfactory receptors in a commercial E.coli cell-free expression system
The ability of peptides to solubilize a wide variety of olfactory receptors was tested by comparing the solubility of all 12 olfactory receptors in 4 peptide detergents and Brij-35 (Figure 2B)
Summary
Olfactory receptors are arguably the most sensitive detectors: they distinguish between thousands of odorants down to parts per billion or trillion. They comprise the largest class of receptors, no molecular structure currently exists, and the molecular basis of olfaction remains an enigma. It is necessary to find an optimal detergent that is capable of keeping them soluble, stable, and functional. Selecting an appropriate detergent is crucial for membrane protein studies, it is a daunting task. Finding an appropriate detergent has become the critical bottleneck for olfactory receptors and other membrane protein studies, and for designing and producing membrane proteins for biotechnological devices
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