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Preparation of functional liposomes with peptide ligands and their binding to cell membranes

  • Published:
Lipids

Abstract

Two novel lipopeptides, which have the peptide ligands [α-melanocyte stimulating hormone (α-MSH)] sequence and repeated [Gly-Arg-Gly-Asp-Se (GRGDS) sequence[, are designed, synthesized by the solid-phase method, and introduced into liposome membranes by the freeze-thaw method. These liposomes bearing the peptide ligands on their surface are expected to bind to cell membranes. We have confirmed that the lipopeptides are introduced into liposome membranes almost quantitatively, while such a high degree of incorporation has not been accomplished in conventional methods. In this respect, the present method is superior to prepare surface-modified liposomes that are applicable to drug carriers and so on. We have also confirmed by using immunoelectron microscopy that the peptide ligands are actually located in an aqueous phase. It has been shown by flow cytometry that the liposome bearing α-MSH peptide ligand binds to B16 cells and the liposome bearing the repeated GRGDS sequence binds to NIH3T3 cells.

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Abbreviations

BSA:

bovine serum albumin

Ch:

cholesterol

CMF:

calcium magesium free

DDS:

drug delivery system

DIPCI:

N,N′-diisopropylcarbodiimide

DMAP:

4-dimethylaminopyridine

DMEM:

Dulbecco's modified Eagle's medium

DMF:

N,N-dimethylformamide

EDT:

1,2-ethanedithiol

EPC:

egg phosphatidylcholine

FITC-PE:

N-fluorescein-5-thiocarbamoyl-1,2-dihexadecanoyl-sn-glycerol-3-phosphoethanolamine

Fmoc:

9-fluorenylmethyloxycarbonyl

GRGDS:

sequence, cell adhesion sequence of fibromectin

HOBt:

1-hydroxybenzotriazole

HPLC:

high-performance liquid chromatography

HRP:

horseradish peroxidase

KLH:

keyhole limpet, hemocyanin

MALDI-TOFMS:

matrix-assisted laser desorption/ionization time of flight mass spectrum

α-MSH:

α-melanocyte-stimulating hormone

NMP:

NMP, N-methylpyrrolidone

NMR:

nuclear magnetic resonance

PBS:

phosphate-buffered saline

TFA:

trifluoroacetic acid

WSCD:

1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride

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Authors and Affiliations

  1. Faculty of Industrial Science and Technology, Science University of Tokyo, 278-8510, Chiba, Japan

    Nobuhiro Yagi & Takeo Konakahara

  2. Biomolecules Department, National Institute of Bioscience and Human-Technology, 1-1 Higashi, 305-8566, Tsukuba, Ibaraki, Japan

    Nobuhiro Yagi, Yoshikatsu Ogawa, Masato Kodaka, Tomoko Okada, Takenori Tomohiro & Hiroaki Okuno

Authors
  1. Nobuhiro Yagi
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  2. Yoshikatsu Ogawa
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  3. Masato Kodaka
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  4. Tomoko Okada
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  5. Takenori Tomohiro
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  6. Takeo Konakahara
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  7. Hiroaki Okuno
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Yagi, N., Ogawa, Y., Kodaka, M. et al. Preparation of functional liposomes with peptide ligands and their binding to cell membranes. Lipids 35, 673–680 (2000). https://doi.org/10.1007/s11745-000-0572-4

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  • DOI: https://doi.org/10.1007/s11745-000-0572-4

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