Time-resolved fluorescence studies of a transmembrane peptide sequence of the dopamine D2 receptor

Valerie L. Williams; Scott H. Courtney; David I. Schuster; Randall B. Murphy

doi:10.1117/12.182759

17 August 1994 Time-resolved fluorescence studies of a transmembrane peptide sequence of the dopamine D₂ receptor

Valerie L. Williams, Scott H. Courtney, David I. Schuster, Randall B. Murphy

Proceedings Volume 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV; (1994) https://doi.org/10.1117/12.182759
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

Highly hydrophobic peptides in small unilamellar vesicles can be used to model membrane-embedded proteins such as the dopamine D₂ receptor. The transmembrane domains of the dopamine D₂ receptor are known to contain residues corresponding to the binding sites for natural receptor ligands. We have developed a model system consisting of a peptide whose sequence was taken from the transmembrane region of the dopamine D₂ receptor and incorporated it into phospholipid bilayers. This polypeptide sequence, NH₂-D-V-L-Y-S-A-F-T-W-L-G-Y-V-N-S-A-V-N-P-I-I-Y-T- T-F-N-V-CO₂H, contains a single tryptophan residue, whose fluorescence properties provides an intrinsic probe of the microenvironment of the peptide within the bilayer. Purification of this highly hydrophobic peptide required the development of a novel alcohol-based reversed-phase HPLC solvent system. The vesicles were produces by cosonication of the peptide with dimyristoylphosphatidylcholine lipid and were characterized by electron microscopy and fluorescence spectroscopy. Time- correlated single photon counting was sued to measure the fluorescence anisotropy of the system as a function of temperature across the lipid phase transition range and as a function of the peptide/lipid ratio.

Citation Download Citation

Valerie L. Williams, Scott H. Courtney, David I. Schuster, and Randall B. Murphy "Time-resolved fluorescence studies of a transmembrane peptide sequence of the dopamine D₂ receptor", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182759

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