Different membrane behaviour and cellular uptake of three basic arginine-rich peptides

Autor: Gérard Chassaing, Eric H. Bent, Nicole Goasdoué, Isabel D. Alves, Isabelle Correia, Claire Lacombe, Olivier Lequin, Astrid Walrant, Sandrine Sagan, Chen-Yu Jiao
Přispěvatelé: Université Pierre et Marie Curie - Paris 6 (UPMC), Service de Biochimie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Bicêtre, Structure et Dynamique des Biomolécules (LBM-E3), Laboratoire des biomolécules (LBM UMR 7203), Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Synthèse, Structure et Fonction de Molécules Bioactives (SSFMB), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Protéines : biochimie structurale et fonctionnelle (PBSF), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC), Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)
Jazyk: angličtina
Rok vydání: 2011
Předmět:
Staphylococcus aureus
Magnetic Resonance Spectroscopy
Antimicrobial peptides
Molecular Sequence Data
Biophysics
Peptide
CHO Cells
Microbial Sensitivity Tests
Calorimetry
010402 general chemistry
Arginine
01 natural sciences
Biochemistry
Cell membrane
Polyarginine
03 medical and health sciences
Cricetulus
Cellular uptake
Cricetinae
medicine
Escherichia coli
Animals
[SDV.BBM]Life Sciences [q-bio]/Biochemistry
Molecular Biology
Amino Acid Sequence
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
chemistry.chemical_classification
[PHYS]Physics [physics]
0303 health sciences
Cell penetrating peptide
Calorimetry
Differential Scanning
Chemistry
[CHIM.ORGA]Chemical Sciences/Organic chemistry
Chinese hamster ovary cell
Vesicle
Circular Dichroism
Cell Membrane
Isothermal titration calorimetry
Cell Biology
NMR
0104 chemical sciences
Anti-Bacterial Agents
Membrane
medicine.anatomical_structure
Cell-penetrating peptide
Peptide lipid interactions
Peptides
Zdroj: Biochimica et Biophysica Acta:Biomembranes
Biochimica et Biophysica Acta:Biomembranes, 2011, 1808 (1), pp.382-393. ⟨10.1016/j.bbamem.2010.09.009⟩
Biochimica et Biophysica Acta:Biomembranes, Elsevier, 2011, 1808 (1), pp.382-393. ⟨10.1016/j.bbamem.2010.09.009⟩
Biochimica et Biophysica Acta-Molecular Cell Research
Biochimica et Biophysica Acta-Molecular Cell Research, Elsevier, 2011, 1808 (1), pp.382-93. ⟨10.1016/j.bbamem.2010.09.009⟩
Biochimica et Biophysica Acta-Molecular Cell Research, 2011, 1808 (1), pp.382-93. ⟨10.1016/j.bbamem.2010.09.009⟩
ISSN: 0005-2736
1879-2642
0167-4889
DOI: 10.1016/j.bbamem.2010.09.009⟩
Popis: International audience; Cell penetrating peptides (CPPs) are peptides displaying the ability to cross cell membranes and transport cargo molecules inside cells. Several uptake mechanisms (endocytic or direct translocation through the membrane) are being considered, but the interaction between the CPP and the cell membrane is certainly a preliminary key point to the entry of the peptide into the cell. In this study, we used three basic peptides: RL9 (RRLLRRLRR-NH(2)), RW9 (RRWWRRWRR-NH(2)) and R9 (RRRRRRRRR-NH(2)). While RW9 and R9 were internalised into wild type Chinese Hamster Ovary cells (CHO) and glycosaminoglycan-deficient CHO cells, at 4°C and 37°C, RL9 was not internalised into CHO cells. To better understand the differences between RW9, R9 and RL9 in terms of uptake, we studied the interaction of these peptides with model lipid membranes. The effect of the three peptides on the thermotropic phase behaviour of a zwitterionic lipid (DMPC) and an anionic lipid (DMPG) was investigated with differential scanning calorimetry (DSC). The presence of negative charges on the lipid headgroups appeared to be essential to trigger the peptide/lipid interaction. RW9 and R9 disturbed the main phase transition of DMPG, whereas RL9 did not induce significant effects. Isothermal titration calorimetry (ITC) allowed us to study the binding of these peptides to large unilamellar vesicles (LUVs). RW9 and R9 proved to have about ten fold more affinity for DSPG LUVs than RL9. With circular dichroism (CD) and NMR spectroscopy, the secondary structure of RL9, RW9 and R9 in aqueous buffer or lipid/detergent conditions was investigated. Additionally, we tested the antimicrobial activity of these peptides against Escherichia coli and Staphylococcus aureus, as CPPs and antimicrobial peptides are known to share several common characteristics. Only RW9 was found to be mildly bacteriostatic against E. coli. These studies helped us to get a better understanding as to why R9 and RW9 are able to cross the cell membrane while RL9 remains bound to the surface without entering the cell.
Databáze: OpenAIRE
Externí odkaz: