title: Tryptophan fluorescence study of the interaction of penetratin peptides with model membranes.
abstract: Penetratin is a 16-amino-acid peptide, derived from the homeodomain of antennapedia, a Drosophila transcription factor, which can be used as a vector for the intracellular delivery of peptides or oligonucleotides. To study the relative importance of the Trp residues in the wild-type penetratin peptide (RQIKIWFQNRRMKWKK) two analogues, the W48F (RQIKIFFQNRRMKWKK) and the W56F (RQI KIWFQNRRMKFKK) variant peptides were synthesized. Binding of the three peptide variants to different lipid vesicles was investigated by fluorescence. Intrinsic Trp fluorescence emission showed a decrease in quantum yield and a blue shift of the maximal emission wavelength upon interaction of the peptides with negatively charged phosphatidylserine, while no changes were recorded with neutral phosphatidylcholine vesicles. Upon binding to phosphatidylcholine vesicles containing 20% (w/w) phosphatidylserine the fluorescence blue shift induced by the W56F-penetratin variant was larger than for the W48F-penetratin. Incorporation of cholesterol into the negatively charged lipid bilayer significantly decreased the binding affinity of the peptides. The Trp mean lifetime of the three peptides decreased upon binding to negatively charged phospholipids, and the Trp residues were shielded from acrylamide and iodide quenching. CD measurements indicated that the peptides are random in buffer, and become alpha helical upon association with negatively charged mixed phosphatidylcholine/phosphatidylserine vesicles, but not with phosphatidylcholine vesicles. These data show that wild-type penetratin and the two analogues interact with negatively charged phospholipids, and that this is accompanied by a conformational change from random to alpha helical structure, and a deeper insertion of W48 compared to W56, into the lipid bilayer.
authors.affiliation: Laboratory for Lipoprotein Chemistry, Ghent University, Belgium.
authors.forename: Bart
authors.initials: B
authors.lastname: Christiaens
authors.forename: Sofie
authors.initials: S
authors.lastname: Symoens
authors.forename: Stefan
authors.initials: S
authors.lastname: Verheyden
authors.forename: Yves
authors.initials: Y
authors.lastname: Engelborghs
authors.forename: Alain
authors.initials: A
authors.lastname: Joliot
authors.forename: Alain
authors.initials: A
authors.lastname: Prochiantz
authors.forename: Joël
authors.initials: J
authors.lastname: Vandekerckhove
authors.forename: Maryvonne
authors.initials: M
authors.lastname: Rosseneu
authors.forename: Berlinda
authors.initials: B
authors.lastname: Vanloo
authors.forename: Stefan
authors.initials: S
authors.lastname: Vanderheyden
mesh_terms: D019943:Amino Acid Substitution; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D057846:Cell-Penetrating Peptides; D002942:Circular Dichroism; D005453:Fluorescence; D008051:Lipid Bilayers / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D014364:Tryptophan / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D008051:Lipid Bilayers; D008563:Membrane Lipids; D010455:Peptides; D014364:Tryptophan; C414409:penetratin