title: Intrinsic tyrosine fluorescence as a tool to study the interaction of the shaker B "ball" peptide with anionic membranes.
abstract: Steady-state and time-resolved fluorescence from the single tyrosine in the inactivating peptide of the Shaker B potassium channel (ShB peptide) and in a noninactivating peptide mutant, ShB-L7E, has been used to characterize their interaction with anionic phospholipid membranes, a model target mimicking features of the inactivation site on the channel protein. Partition coefficients derived from steady-state anisotropy indicate that both peptides show a high affinity for anionic vesicles, being higher in ShB than in ShB-L7E. Moreover, differential quenching by lipophilic spin-labeled probes and fluorescence energy transfer using trans-parinaric acid as the acceptor confirm that the ShB peptide inserts deep into the membrane, while the ShB-L7E peptide remains near the membrane surface. The rotational mobility of tyrosine in membrane-embedded ShB, examined from the decay of fluorescence anisotropy, can be described by two different rotational correlation times and a residual constant value. The short correlation time corresponds to fast rotation reporting on local tyrosine mobility. The long rotational correlation time and the high residual anisotropy suggest that the ShB peptide diffuses in a viscous and anisotropic medium compatible with the aliphatic region of a lipid bilayer and support the hypothesis that the peptide inserts into it as a monomer, to configure an intramolecular beta-hairpin structure. Assuming that this hairpin structure behaves like a rigid body, we have estimated its dimensions and rotational dynamics, and a model for the peptide inserted into the bilayer has been proposed.
authors.affiliation: Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, E-03202 Elche (Alicante), Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Poveda
authors.forename: Manuel
authors.initials: M
authors.lastname: Prieto
authors.forename: José A
authors.initials: JA
authors.lastname: Encinar
authors.forename: José M
authors.initials: JM
authors.lastname: González-Ros
authors.forename: C Reyes
authors.initials: CR
authors.lastname: Mateo
mesh_terms: D000595:Amino Acid Sequence; D005454:Fluorescence Polarization; D031541:Fluorescence Resonance Energy Transfer; D008567:Membranes, Artificial; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism*; D015221:Potassium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D011789:Quantum Theory; D051637:Shaker Superfamily of Potassium Channels; D014443:Tyrosine / Q000737:chemistry*
chemical_list: D008567:Membranes, Artificial; D010743:Phospholipids; D015221:Potassium Channels; D051637:Shaker Superfamily of Potassium Channels; D014443:Tyrosine