title: The orientation and dynamics of substance P in lipid environments.
abstract: The membrane-associated conformation of substance P (RPKPQQFFGLM-NH2) has been previously proposed to be the NK1-receptor-active conformation. In this work, NMR methods are applied to explore the orientation and dynamics of substance P at lipid surfaces for which the peptide's three-dimensional structure had been previously determined. Here the presence of dodecylphosphocholine (DPC) or sodium dodecylsulfate (SDS) micelles has been found to cause sequence specific changes in the acid- and base-catalyzed amide proton exchange rates relative to the solution state values. On binding of substance P to SDS micelles, the FFG portion showed the largest decreases in the base-catalyzed amide exchange rates. Similar sequence-specific changes in substance P are observed in the presence of DPC micelles, albeit at much weaker levels due to fast exchange between free and bound forms of the peptide. These differences are attributed to the location of the amide protons either in the surface double layer (via electrostatic effect) or inserted into the polar head group region of the micelles (via low dielectric). The sequence-specific effects of micelle association were also observed in the homonuclear nonselective spin-lattice relaxation time; these, in combination with spin-spin relaxation times, were used to calculate correlation times for the backbone amide protons. These data combined with paramagnetic broadening observations on peptide protons in the presence of spin-labeled lipids yield a detailed model of the interaction of substance P with lipid surfaces.
authors.affiliation: The Beckman Research Institute of the City of Hope, Duarte, California 91010-0269, USA. dak@ernst.coh.org
authors.forename: D A
authors.initials: DA
authors.lastname: Keire
authors.forename: M
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000595:Amino Acid Sequence; D006863:Hydrogen-Ion Concentration; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008433:Mathematics; D008823:Micelles; D010767:Phosphorylcholine / Q000031:analogs & derivatives; D011487:Protein Conformation; D012967:Sodium Dodecyl Sulfate; D013113:Spin Labels; D055672:Static Electricity; D013373:Substance P / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008055:Lipids; D008823:Micelles; D013113:Spin Labels; D010767:Phosphorylcholine; D013373:Substance P; D012967:Sodium Dodecyl Sulfate; C028810:dodecylphosphocholine