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The Journal of Membrane Biology

, Volume 251, Issue 4, pp 581–592 | Cite as

Using Liprotides to Deliver Cholesterol to the Plasma Membrane

  • Henriette S. Frislev
  • Janni Nielsen
  • Jesper Nylandsted
  • Daniel Otzen
Article
  • 119 Downloads

Abstract

Cholesterol (chol) is important in all mammalian cells as a modulator of membrane fluidity. However, its low solubility is a challenge for controlled delivery to membranes. Here we introduce a new tool to deliver chol to membranes, namely, liprotides, i.e., protein–lipid complexes composed of a fatty acid core decorated with partially denatured protein. We focus on liprotides prepared by incubating Ca2+-depleted α-lactalbumin with oleic acid (OA) for 1 h at 20 °C (lip20) or 80 °C (lip80). The binding and membrane delivery properties of liprotides is compared to the widely chol transporter methyl-β-cyclodextrin (mBCD). Both lip20 and lip80 increase the solubility of chol ~ 50% more than mBCD and deliver chol to membranes with comparable efficiency. Although OA is cytotoxic at high concentrations, its effects are counterbalanced by chol. Further, cytotoxicity is strongly reduced when OA is replaced by cis-palmitoleic acid or cis-vaccenic acid. This makes liprotides good tools to deliver chol to membranes and cells.

Keywords

Cholesterol Liprotides Membrane Cytotoxicity 

Abbreviations

aLA

α-Lactalbumin

chol

Cholesterol

cPA

cis-Palmitoleic acid

cVA

cis-Vaccenic acid

DOPC

1,2-Dioleoylphosphatidylcholine

DOPG

1,2-Dioleoylphosphatidylglycerol

HAMLET

Human α-lactalbumin made lethal to tumor cells

Liprotides

Complexes between lipids and partially denatured proteins

lip20

Liprotide prepared at 20 °C

lip80

Liprotide prepared at 80 °C, lip:chol, liprotide in complex with chol

mBCD

Methyl-β-cyclodextrin

mBCD:chol

mBCD in complex with chol

OA

Oleic acid

QCM-D

Quartz crystal microbalance with dissipation

RP-HPLC

Reversed-phase high-performance liquid chromatography

RT

Room temperature

TFA

Trifluoroacetic acid

Notes

Acknowledgements

This work is supported by a grant from the Danish Research Council | Technology and Production (DFF-4005-00479) to D.E.O. and H.S.F. We are grateful to Camilla Bertel Andersen for help with analysis of chol.

Compliance with Ethical Standards

Conflict of interest

The authors declare no competing financial interests.

Supplementary material

232_2018_34_MOESM1_ESM.docx (11.1 mb)
Supplementary Information includes figures depicting the amount of transfer of content to vesicles and the membrane destabilizing effect of liprotides (DOCX 11362 KB)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Interdisciplinary Nanoscience Center (iNANO), Department of Molecular Biology and GeneticsAarhus UniversityAarhus CDenmark
  2. 2.Membrane Integrity Group, Cell Death and Metabolism UnitDanish Cancer Society Research CenterCopenhagenDenmark

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