title: Anchorage of synthetic peptides onto liposomes via hydrazone and alpha-oxo hydrazone bonds. preliminary functional investigations.
abstract: Synthetic peptidoliposomes have been designed and prepared according to a chemoselective ligation. Two aldehyde-functionalized lipidic anchors were synthesized and incorporated into the lipidic bilayers of unilamellar liposomes during their preparation. Complementary hydrazino acetyl peptides were synthesized on the solid phase using N,N',N'-tri(tert-butyloxycarbonyl)-hydrazino acetic acid and further coupled to the aldehyde groups displayed at the surface of the vesicles. Coupling yields were measured by amino acid hydrolysis following total acid hydrolysis. The ligation methodology proved superior to the simple insertion of lipopeptides, which was performed for comparison in terms of yields, implementation, and reproducibility. To check whether the grafted-peptides were accessible and functional, cytoplasmic sequences of LAMP protein (lysosomal associated membrane protein), which is involved in intracellular membrane trafficking, have been selected. Using this model, we demonstrated in vitro the specific interaction of the synthetic LAMP-peptidoliposomes with the cytoplasmic adaptor protein AP-3, a result that contributes to the understanding of protein sorting in cells. Thus, these results clearly indicate the usefulness of such peptidoliposomes, easily prepared by hydrazone chemoselective ligation, as a tool for biological investigation.
authors.affiliation: UMR 8525 CNRS, Institut Pasteur de Lille et Université de Lille 2, France, FRE 2377 CNRS, Institut de Biologie de Lille, 1 rue du Pr. Calmette, 59021 Lille, France. lbourel@pharma.univ-lille2.fr
authors.forename: Line
authors.initials: L
authors.lastname: Bourel-Bonnet
authors.forename: Eve-Isabelle
authors.initials: EI
authors.lastname: Pécheur
authors.forename: Cyrille
authors.initials: C
authors.lastname: Grandjean
authors.forename: Annick
authors.initials: A
authors.lastname: Blanpain
authors.forename: Thorsten
authors.initials: T
authors.lastname: Baust
authors.forename: Oleg
authors.initials: O
authors.lastname: Melnyk
authors.forename: Bernard
authors.initials: B
authors.lastname: Hoflack
authors.forename: Hélène
authors.initials: H
authors.lastname: Gras-Masse
mesh_terms: D033963:Adaptor Protein Complex 3; D033981:Adaptor Protein Complex delta Subunits; D015703:Antigens, CD; D020650:Combinatorial Chemistry Techniques; D006835:Hydrazones / Q000737:chemistry*; D008081:Liposomes / Q000138:chemical synthesis*; D051907:Lysosomal Membrane Proteins; D015335:Molecular Probes / Q000138:chemical synthesis*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D021381:Protein Transport; D014157:Transcription Factors / Q000378:metabolism
chemical_list: C491904:AP3D1 protein, human; D033963:Adaptor Protein Complex 3; D033981:Adaptor Protein Complex delta Subunits; D015703:Antigens, CD; D006835:Hydrazones; D008081:Liposomes; D051907:Lysosomal Membrane Proteins; D015335:Molecular Probes; D010455:Peptides; D014157:Transcription Factors