Membrane affinity and fluorescent labelling

comparative study of monolayer interaction, cellular uptake and cytotoxicity profile of carboxyfluorescein-conjugated cationic peptides

E. Kiss, Gergő Gyulai, Edit Pári, Kata Horváti, Sz. Bősze

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Fluorescent labelling is a common approach to reveal the molecular details of cellular uptake, internalisation, transport, distribution processes in biological systems. The conjugation with a fluorescent moiety might affect relevant physico-chemical and in vitro transport properties of the bioactive component. A representative set of seven cationic peptides—including cell-penetrating peptides as well as antimicrobial peptides and synthetic derivatives—was selected for our comparative study. Membrane affinity of the peptides and their 5(6)-carboxyfluorescein (Cf) derivatives was determined quantitatively and compared applying Langmuir monolayer of zwitterionic (DPPC) and negatively charged (DPPC + DPPG) lipids as cell membrane models. The interaction with neutral lipid layer is mainly governed by the overall hydrophobicity of the molecule which is remarkably increased by Cf-conjugation for the most hydrophobic Magainin, Melittin and Transportan. A significantly enhanced membrane affinity was detected in negatively charged lipid model monolayer for all of the peptides since the combination of electrostatic and hydrophobic interaction is active in that case. The Cf-conjugation improved the penetration ability of Penetratin and Dhvar4 suggesting that both the highly charged character (Z/n) and the increased hydrophobicity by Cf-conjugation present important contribution to membrane interaction. This effect might also responsible for the observed high in vitro internalisation rate of Penetratin and Dhvar4, while according to in vitro studies they did not cause damage of cell membrane. From the experiments with the given seven cationic peptides, it can be concluded that the Cf-conjugation alters the degree of membrane interaction of such peptides which are moderately hydrophobic and highly charged.

Original languageEnglish
JournalAmino Acids
DOIs
Publication statusAccepted/In press - Jan 1 2018

Fingerprint

Cytotoxicity
Labeling
Monolayers
Membranes
Peptides
Hydrophobic and Hydrophilic Interactions
Cell membranes
Hydrophobicity
Lipids
Magainins
Cell-Penetrating Peptides
Melitten
Biological Phenomena
Biological systems
Membrane Lipids
Static Electricity
Transport properties
Electrostatics
Cell Membrane
6-carboxyfluorescein

Keywords

  • Cell-penetrating peptides
  • Cellular uptake
  • Fluorescent labelling
  • Lipid monolayer
  • Membrane affinity
  • Penetration

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

@article{9adfa00a9f524c57a0eca70a319b12b0,
title = "Membrane affinity and fluorescent labelling: comparative study of monolayer interaction, cellular uptake and cytotoxicity profile of carboxyfluorescein-conjugated cationic peptides",
abstract = "Fluorescent labelling is a common approach to reveal the molecular details of cellular uptake, internalisation, transport, distribution processes in biological systems. The conjugation with a fluorescent moiety might affect relevant physico-chemical and in vitro transport properties of the bioactive component. A representative set of seven cationic peptides—including cell-penetrating peptides as well as antimicrobial peptides and synthetic derivatives—was selected for our comparative study. Membrane affinity of the peptides and their 5(6)-carboxyfluorescein (Cf) derivatives was determined quantitatively and compared applying Langmuir monolayer of zwitterionic (DPPC) and negatively charged (DPPC + DPPG) lipids as cell membrane models. The interaction with neutral lipid layer is mainly governed by the overall hydrophobicity of the molecule which is remarkably increased by Cf-conjugation for the most hydrophobic Magainin, Melittin and Transportan. A significantly enhanced membrane affinity was detected in negatively charged lipid model monolayer for all of the peptides since the combination of electrostatic and hydrophobic interaction is active in that case. The Cf-conjugation improved the penetration ability of Penetratin and Dhvar4 suggesting that both the highly charged character (Z/n) and the increased hydrophobicity by Cf-conjugation present important contribution to membrane interaction. This effect might also responsible for the observed high in vitro internalisation rate of Penetratin and Dhvar4, while according to in vitro studies they did not cause damage of cell membrane. From the experiments with the given seven cationic peptides, it can be concluded that the Cf-conjugation alters the degree of membrane interaction of such peptides which are moderately hydrophobic and highly charged.",
keywords = "Cell-penetrating peptides, Cellular uptake, Fluorescent labelling, Lipid monolayer, Membrane affinity, Penetration",
author = "E. Kiss and Gergő Gyulai and Edit P{\'a}ri and Kata Horv{\'a}ti and Sz. Bősze",
year = "2018",
month = "1",
day = "1",
doi = "10.1007/s00726-018-2630-7",
language = "English",
journal = "Amino Acids",
issn = "0939-4451",
publisher = "Springer Wien",

}

TY - JOUR

T1 - Membrane affinity and fluorescent labelling

T2 - comparative study of monolayer interaction, cellular uptake and cytotoxicity profile of carboxyfluorescein-conjugated cationic peptides

AU - Kiss, E.

AU - Gyulai, Gergő

AU - Pári, Edit

AU - Horváti, Kata

AU - Bősze, Sz.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Fluorescent labelling is a common approach to reveal the molecular details of cellular uptake, internalisation, transport, distribution processes in biological systems. The conjugation with a fluorescent moiety might affect relevant physico-chemical and in vitro transport properties of the bioactive component. A representative set of seven cationic peptides—including cell-penetrating peptides as well as antimicrobial peptides and synthetic derivatives—was selected for our comparative study. Membrane affinity of the peptides and their 5(6)-carboxyfluorescein (Cf) derivatives was determined quantitatively and compared applying Langmuir monolayer of zwitterionic (DPPC) and negatively charged (DPPC + DPPG) lipids as cell membrane models. The interaction with neutral lipid layer is mainly governed by the overall hydrophobicity of the molecule which is remarkably increased by Cf-conjugation for the most hydrophobic Magainin, Melittin and Transportan. A significantly enhanced membrane affinity was detected in negatively charged lipid model monolayer for all of the peptides since the combination of electrostatic and hydrophobic interaction is active in that case. The Cf-conjugation improved the penetration ability of Penetratin and Dhvar4 suggesting that both the highly charged character (Z/n) and the increased hydrophobicity by Cf-conjugation present important contribution to membrane interaction. This effect might also responsible for the observed high in vitro internalisation rate of Penetratin and Dhvar4, while according to in vitro studies they did not cause damage of cell membrane. From the experiments with the given seven cationic peptides, it can be concluded that the Cf-conjugation alters the degree of membrane interaction of such peptides which are moderately hydrophobic and highly charged.

AB - Fluorescent labelling is a common approach to reveal the molecular details of cellular uptake, internalisation, transport, distribution processes in biological systems. The conjugation with a fluorescent moiety might affect relevant physico-chemical and in vitro transport properties of the bioactive component. A representative set of seven cationic peptides—including cell-penetrating peptides as well as antimicrobial peptides and synthetic derivatives—was selected for our comparative study. Membrane affinity of the peptides and their 5(6)-carboxyfluorescein (Cf) derivatives was determined quantitatively and compared applying Langmuir monolayer of zwitterionic (DPPC) and negatively charged (DPPC + DPPG) lipids as cell membrane models. The interaction with neutral lipid layer is mainly governed by the overall hydrophobicity of the molecule which is remarkably increased by Cf-conjugation for the most hydrophobic Magainin, Melittin and Transportan. A significantly enhanced membrane affinity was detected in negatively charged lipid model monolayer for all of the peptides since the combination of electrostatic and hydrophobic interaction is active in that case. The Cf-conjugation improved the penetration ability of Penetratin and Dhvar4 suggesting that both the highly charged character (Z/n) and the increased hydrophobicity by Cf-conjugation present important contribution to membrane interaction. This effect might also responsible for the observed high in vitro internalisation rate of Penetratin and Dhvar4, while according to in vitro studies they did not cause damage of cell membrane. From the experiments with the given seven cationic peptides, it can be concluded that the Cf-conjugation alters the degree of membrane interaction of such peptides which are moderately hydrophobic and highly charged.

KW - Cell-penetrating peptides

KW - Cellular uptake

KW - Fluorescent labelling

KW - Lipid monolayer

KW - Membrane affinity

KW - Penetration

UR - http://www.scopus.com/inward/record.url?scp=85051453798&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85051453798&partnerID=8YFLogxK

U2 - 10.1007/s00726-018-2630-7

DO - 10.1007/s00726-018-2630-7

M3 - Article

JO - Amino Acids

JF - Amino Acids

SN - 0939-4451

ER -